Immunoregulatory agents

ABSTRACT

Compounds that modulate the oxidoreductase enzyme indoleamine 2,3-dioxygenase, and compositions containing the compounds, are described herein. The use of such compounds and compositions for the treatment and/or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by indoleamine 2,3-dioxygenase is also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application Ser.No. 62/075,678, filed Nov. 5, 2014, the entire content of which isincorporated herein by reference.

BACKGROUND OF THE INVENTION

Indoleamine 2,3-dioxygenase (IDO; also known as IDO1) is an IFN-γ targetgene that plays a role in immunomodulation. IDO is an oxidoreductase andone of two enzymes that catalyze the first and rate-limiting step in theconversion of tryptophan to N-formyl-kynurenine. It exists as a 41 kDmonomer that is found in several cell populations, including immunecells, endothelial cells, and fibroblasts. IDO is relativelywell-conserved between species, with mouse and human sharing 63%sequence identity at the amino acid level. Data derived from its crystalstructure and site-directed mutagenesis show that both substrate bindingand the relationship between the substrate and iron-bound dioxygenaseare necessary for activity. A homolog to IDO (IDO2) has been identifiedthat shares 44% amino acid sequence homology with IDO, but its functionis largely distinct from that of IDO. (See, e.g., Serafini, P. et al.,Semin. Cancer Biol., 16(1):53-65 (February 2006) and Ball, H. J. et al.,Gene, 396(1):203-213 (Jul. 1, 2007)).

IDO plays a major role in immune regulation, and its immunosuppressivefunction manifests in several manners. Importantly, IDO regulatesimmunity at the T cell level, and a nexus exists between IDO andcytokine production. In addition, tumors frequently manipulate immunefunction by upregulation of IDO. Thus, modulation of IDO can have atherapeutic impact on a number of diseases, disorders and conditions.

A pathophysiological link exists between IDO and cancer. Disruption ofimmune homeostasis is intimately involved with tumor growth andprogression, and the production of IDO in the tumor microenvironmentappears to aid in tumor growth and metastasis. Moreover, increasedlevels of IDO activity are associated with a variety of different tumors(Brandacher, G. et al., Clin. Cancer Res., 12(4):1144-1151 (Feb. 15,2006)).

Treatment of cancer commonly entails surgical resection followed bychemotherapy and radiotherapy. The standard treatment regimens showhighly variable degrees of long-term success because of the ability oftumor cells to essentially escape by regenerating primary tumor growthand, often more importantly, seeding distant metastasis. Recent advancesin the treatment of cancer and cancer-related diseases, disorders andconditions comprise the use of combination therapy incorporatingimmunotherapy with more traditional chemotherapy and radiotherapy. Undermost scenarios, immunotherapy is associated with less toxicity thantraditional chemotherapy because it utilizes the patient's own immunesystem to identify and eliminate tumor cells.

In addition to cancer, IDO has been implicated in, among otherconditions, immunosuppression, chronic infections, and autoimmunediseases or disorders (e.g., rheumatoid arthritis). Thus, suppression oftryptophan degradation by inhibition of IDO activity has tremendoustherapeutic value. Moreover, inhibitors of IDO can be used to enhance Tcell activation when the T cells are suppressed by pregnancy,malignancy, or a virus (e.g., HIV). Although their roles are not as welldefined, IDO inhibitors may also find use in the treatment of patientswith neurological or neuropsychiatric diseases or disorders (e.g.,depression).

Small molecule inhibitors of IDO have been developed to treat or preventIDO-related diseases. For example, the IDO inhibitors1-methyl-DL-tryptophan; p-(3-benzofuranyl)-DL-alanine;p-[3-benzo(b)thienyl]-DL-alanine; and 6-nitro-L-tryptophan have beenused to modulate T cell-mediated immunity by altering localextracellular concentrations of tryptophan and tryptophan metabolites(WO 99/29310). Compounds having IDO inhibitory activity are furtherreported in PCT Publication No. WO 2004/094409.

In view of the role played by indoleamine 2,3-dioxygenase in a diversearray of diseases, disorders and conditions, and the limitations (e.g.,efficacy) of current IDO inhibitors, new IDO modulators, andcompositions and methods associated therewith, are needed.

BRIEF SUMMARY OF THE INVENTION

The present invention relates to compounds that modulate theoxidoreductase enzyme indoleamine 2,3-dioxygenase (IDO), andcompositions (e.g., pharmaceutical compositions) comprising thecompounds. Such compounds, including methods of their synthesis, andcompositions are described in detail below.

The present invention also relates to the use of such compounds andcompositions for the treatment and/or prevention of a diverse array ofdiseases, disorders and conditions mediated, in whole or in part, byIDO. Such diseases, disorders and conditions are described in detailelsewhere herein. Unless otherwise indicated, when uses of the compoundsof the present invention are described herein, it is to be understoodthat such compounds may be in the form of a composition (e.g., apharmaceutical composition).

As discussed hereafter, although the compounds of the present inventionare believed to effect their activity by inhibition of IDO, a preciseunderstanding of the compounds' underlying mechanism of action is notrequired to practice the invention. It is envisaged that the compoundsmay alternatively effect their activity through inhibition oftryptophan-2,3-dioxygenase (TDO) activity. It is also envisaged that thecompounds may effect their activity through inhibition of both IDO andTDO function. Although the compounds of the invention are generallyreferred to herein as IDO inhibitors, it is to be understood that theterm “IDO inhibitors” encompasses compounds that act individuallythrough inhibition of TDO or IDO, and/or compounds that act throughinhibition of both IDO and TDO.

In one aspect, the present invention provides compounds represented byformula

or a pharmaceutically acceptable salt, hydrate or solvate thereof,wherein,

-   the subscript n is 1 or 0;-   A is —C(O)—, —NH—, —SO₂—, —CH₂—, or —CHR³—;-   B is a bond, —C(O)—, —NH—, —CH₂—, or —CHR³—;-   T is a bond, —CH₂—, —NH—, —O—, —OCH₂—, —C(O)CH₂—, or —CR³R⁴—;    -   wherein when A is —NH— and B is —C(O)—, then T is other than        —C(R³)(R⁴)—; D is N or C(R⁵);-   E is N or C(R⁶);-   V is a bond, —O—, or —C(R^(5a))₂;-   G is an optionally substituted aryl, optionally substituted    heteroaryl, or an optionally substituted 9- or 10-membered fused    bicyclic heteroaryl;-   J¹ is CH, N or C(R²), when R² is attached to the ring vertex    identified as J¹;-   R¹ and R² are independently hydrogen, halogen, optionally    substituted C₁-C₄ haloalkyl, optionally substituted C₃-C₆    cycloalkyl, optionally substituted 3- to 6-membered    cycloheteroalkyl, optionally substituted phenyl, optionally    substituted heteroaryl, optionally substituted C₁-C₄ alkyl,    optionally substituted C₁-C₄ alkoxy, CN, SO₂NH₂, NHSO₂CH₃, NHSO₂CF₃,    OCF₃, SO₂CH₃, SO₂CF₃, or CONH₂, and when R¹ and R² are on adjacent    vertices of a phenyl ring they may be joined together to form a 5-    or 6-membered cycloheteroalkyl ring having one or two ring vertices    independently selected from O, N and S, wherein said    cycloheteroalkyl ring is optionally substituted with from one to    three members selected from fluoro and C₁-C₃ alkyl;-   R³ and R⁴ are independently hydrogen, optionally substituted C₁-C₆    alkyl, optionally substituted C₁-C₆ haloalkyl, fluorine, OH, CN,    CO₂H, C(O)NH₂, N(R^(5a))₂, optionally substituted —O—C₁-C₆ alkyl,    —(CR⁵R⁵)_(m)—OH, —(CR⁵R⁵)_(m)—CO₂H, —(CR⁵R⁵)_(m)—C(O)NH₂,    —(CR⁵R⁵)_(m)—C(O)NHR^(5a), —(CR⁵R⁵)_(m)N(R^(5a))₂,    —NH(CR⁵R⁵)_(m)CO₂H or —NH(CR⁵R⁵)_(m)—C(O)NH₂;-   each R⁵ is independently H, F, OH, optionally substituted C₁-C₆    alkyl or optionally substituted —O—C₁-C₆ alkyl;-   each R^(5a) is independently H, or optionally substituted C₁-C₆    alkyl;-   R⁶ is H, OH, F, optionally substituted C₁-C₆ alkyl, optionally    substituted —O—C₁-C₆ alkyl, or —N(R^(5a))₂;-   and each m is independently 1, 2, or 3.

In yet another aspect, the present invention provides compositions inwhich compounds of formula (I), are combined with one or morepharmaceutically acceptable excipients.

In some embodiments, the present invention contemplates methods fortreating or preventing cancer in a subject (e.g., a human) comprisingadministering to the subject a therapeutically effective amount of atleast one IDO inhibitor described herein. The present invention includesmethods of treating or preventing a cancer in a subject by administeringto the subject an IDO inhibitor in an amount effective to reverse orstop the progression of IDO-mediated immunosuppression. In someembodiments, the IDO-mediated immunosuppression is mediated by anantigen-presenting cell (APC).

Examples of the cancers that may be treated using the compounds andcompositions described herein include, but are not limited to: cancersof the prostate, colorectum, pancreas, cervix, stomach, endometrium,brain, liver, bladder, ovary, testis, head, neck, skin (includingmelanoma and basal carcinoma), mesothelial lining, white blood cell(including lymphoma and leukemia) esophagus, breast, muscle, connectivetissue, lung (including small-cell lung carcinoma and non-small-cellcarcinoma), adrenal gland, thyroid, kidney, or bone; glioblastoma,mesothelioma, renal cell carcinoma, gastric carcinoma, sarcoma,choriocarcinoma, cutaneous basocellular carcinoma, and testicularseminoma. In some embodiments of the present invention, the cancer ismelanoma, colon cancer, pancreatic cancer, breast cancer, prostatecancer, lung cancer, leukemia, a brain tumor, lymphoma, sarcoma, ovariancancer, head and neck cancer, cervical cancer, or Kaposi's sarcoma.Cancers that are candidates for treatment with the compounds andcompositions of the present invention are discussed further hereafter.

The present invention contemplates methods of treating a subjectreceiving a bone marrow transplant or peripheral blood stem celltransplant by administering a therapeutically effective amount of an IDOinhibitor sufficient to increase the delayed-type hypersensitivityreaction to tumor antigen, delay the time-to-relapse of post-transplantmalignancy, increase relapse-free survival time post-transplant, and/orincrease long-term post-transplant survival.

In certain embodiments, the present invention contemplates methods fortreating or preventing an infective disorder (e.g., a viral infection)in a subject (e.g., a human) comprising administering to the subject atherapeutically effective amount of at least one IDO inhibitor (e.g., anovel inhibitor of the instant invention). In some embodiments, theinfective disorder is a viral infection (e.g., a chronic viralinfection), a bacterial infection, or a parasitic infection. In certainembodiments, the viral infection is human immunodeficiency virus orcytomegalovirus. In other embodiments, the bacterial infection is aMycobacterium infection (e.g., Mycobacterium leprae or Mycobacteriumtuberculosis). In still other embodiments, the parasitic infection isLeishmania donovani, Leishmania tropica, Leishmania major, Leishmaniaaethiopica, Leishmania mexicana, Plasmodium falciparum, Plasmodiumvivax, Plasmodium ovale, or Plasmodium malariae. In further embodiments,the infective disorder is a fungal infection.

In still other embodiments, the present invention contemplates methodsfor treating or preventing an immune-related disease, disorder orcondition in a subject (e.g., a human), comprising administering to thesubject a therapeutically effective amount of at least one IDO inhibitor(e.g., preferably a novel inhibitor of the instant invention). Examplesof immune-related diseases, disorders and conditions are describedhereafter.

Other diseases, disorders and conditions that may be treated orprevented, in whole or in part, by modulation of IDO activity arecandidate indications for the IDO inhibitor compounds that are describedherein.

The present invention further contemplates the use of the IDO inhibitorsdescribed herein in combination with one or more additional agents. Theone or more additional agents may have some IDO modulating activityand/or they may function through distinct mechanisms of action. In someembodiments, such agents comprise radiation (e.g., localized radiationtherapy or total body radiation therapy) and/or other treatmentmodalities of a non-pharmacological nature. When combination therapy isutilized, the IDO inhibitor(s) and the one additional agent(s) may be inthe form of a single composition or multiple compositions, and thetreatment modalities may be administered concurrently, sequentially, orthrough some other regimen. By way of example, the present inventioncontemplates a treatment regimen wherein a radiation phase is followedby a chemotherapeutic phase. The combination therapy may have anadditive or synergistic effect. Other benefits of combination therapyare described hereafter.

In some embodiments, the present invention further comprises the use ofthe IDO inhibitors described herein in combination with bone marrowtransplantation, peripheral blood stem cell transplantation, or othertypes of transplantation therapy.

In particular embodiments, the present invention contemplates the use ofthe inhibitors of IDO function described herein in combination withimmune checkpoint inhibitors. The blockade of immune checkpoints, whichresults in the amplification of antigen-specific T cell responses, hasbeen shown to be a promising approach in human cancer therapeutics.Examples of immune checkpoints (ligands and receptors), some of whichare selectively upregulated in various types of tumor cells, that arecandidates for blockade include PD1 (programmed cell death protein 1);PDL1 (PD1 ligand); BTLA (B and T lymphocyte attenuator); CTLA4(cytotoxic T-lymphocyte associated antigen 4); TIM3 (T-cell membraneprotein 3); LAG3 (lymphocyte activation gene 3); A2aR (adenosine A2areceptor A2aR); and Killer Inhibitory Receptors. Immune checkpointinhibitors, and combination therapy therewith, are discussed in detailelsewhere herein.

In other embodiments, the present invention provides methods fortreating cancer in a subject, comprising administering to the subject atherapeutically effective amount of at least one IDO inhibitor and atleast one chemotherapeutic agent, such agents including, but not limitedto alkylating agents (e.g., nitrogen mustards such as chlorambucil,cyclophosphamide, isofamide, mechlorethamine, melphalan, and uracilmustard; aziridines such as thiotepa; methanesulphonate esters such asbusulfan; nucleoside analogs (e.g., gemcitabine); nitroso ureas such ascarmustine, lomustine, and streptozocin; topoisomerase 1 inhibitors(e.g., irinotecan); platinum complexes such as cisplatin andcarboplatin; bioreductive alkylators such as mitomycin, procarbazine,dacarbazine and altretamine); DNA strand-breakage agents (e.g.,bleomycin); topoisomerase II inhibitors (e.g., amsacrine, dactinomycin,daunorubicin, idarubicin, mitoxantrone, doxorubicin, etoposide, andteniposide); DNA minor groove binding agents (e.g., plicamydin);antimetabolites (e.g., folate antagonists such as methotrexate andtrimetrexate; pyrimidine antagonists such as fluorouracil,fluorodeoxyuridine, CB3717, azacitidine, cytarabine, and floxuridine;purine antagonists such as mercaptopurine, 6-thioguanine, fludarabine,pentostatin; asparginase; and ribonucleotide reductase inhibitors suchas hydroxyurea); tubulin interactive agents (e.g., vincristine,estramustine, vinblastine, docetaxol, epothilone derivatives, andpaclitaxel); hormonal agents (e.g., estrogens; conjugated estrogens;ethinyl estradiol; diethylstilbesterol; chlortrianisen; idenestrol;progestins such as hydroxyprogesterone caproate, medroxyprogesterone,and megestrol; and androgens such as testosterone, testosteronepropionate, fluoxymesterone, and methyltestosterone); adrenalcorticosteroids (e.g., prednisone, dexamethasone, methylprednisolone,and prednisolone); leutinizing hormone releasing agents orgonadotropin-releasing hormone antagonists (e.g., leuprolide acetate andgoserelin acetate); and antihormonal antigens (e.g., tamoxifen,antiandrogen agents such as flutamide; and antiadrenal agents such asmitotane and aminoglutethimide). The present invention also contemplatesthe use of the IDO inhibitors in combination with other agents known inthe art (e.g., arsenic trioxide) and other chemotherapeutic agentsdeveloped in the future.

In some embodiments drawn to methods of treating cancer, theadministration of a therapeutically effective amount of an IDO inhibitorin combination with at least one chemotherapeutic agent results in acancer survival rate greater than the cancer survival rate observed byadministering either alone. In further embodiments drawn to methods oftreating cancer, the administration of a therapeutically effectiveamount of an IDO inhibitor in combination with at least onechemotherapeutic agent results in a reduction of tumor size or a slowingof tumor growth greater than reduction of the tumor size or tumor growthobserved by administration of one agent alone.

In further embodiments, the present invention contemplates methods fortreating or preventing cancer in a subject, comprising administering tothe subject a therapeutically effective amount of at least one IDOinhibitor and at least one signal transduction inhibitor (STI). In aparticular embodiment, the at least one STI is selected from the groupconsisting of bcr/abl kinase inhibitors, epidermal growth factor (EGF)receptor inhibitors, her-2/neu receptor inhibitors, and farnesyltransferase inhibitors (FTIs). Other candidate STI agents are set forthelsewhere herein.

The present invention also contemplates methods of augmenting therejection of tumor cells in a subject comprising administering an IDOinhibitor in conjunction with at least one chemotherapeutic agent and/orradiation therapy, wherein the resulting rejection of tumor cells isgreater than that obtained by administering either the IDO inhibitor,the chemotherapeutic agent or the radiation therapy alone.

In further embodiments, the present invention provides methods fortreating cancer in a subject, comprising administering to the subject atherapeutically effective amount of at least one IDO inhibitor and atleast one immunomodulator other than an IDO inhibitor. In particularembodiments, the at least one immunomodulator is selected from the groupconsisting of CD40L, B7, B7RP1, ant-CD40, anti-CD38, anti-ICOS, 4-IBBligand, dendritic cell cancer vaccine, IL2, IL12, ELC/CCL19, SLC/CCL21,MCP-1, IL-4, IL-18, TNF, IL-15, MDC, IFN-α/-β, M-CSF, IL-3, GM-CSF,IL-13, and anti-IL-10. Other candidate immunomodulator agents are setforth elsewhere herein.

The present invention contemplates embodiments comprising methods fortreating or preventing an infective disorder (e.g., a viral infection)in a subject (e.g., a human) comprising administering to the subject atherapeutically effective amount of at least one IDO inhibitor and atherapeutically effective amount of an anti-infective agent(s)

In some embodiments of the present invention, the additional therapeuticagent is a cytokine, including, for example, granulocyte-macrophagecolony stimulating factor (GM-CSF) or flt3-ligand. The present inventionalso contemplates methods for treating or preventing a viral infection(e.g., a chronic viral infection) including, but not limited to,hepatitis C virus (HCV), human papilloma virus (HPV), cytomegalovirus(CMV), Epstein-Barr virus (EBV), varicella zoster virus, coxsackievirus, and human immunodeficiency virus (HIV). The use of the IDOinhibitors described herein to treat (either alone or as a component ofcombination therapy) infection is discussed further hereafter.

In additional embodiments, treatment of an infective disorder iseffected through the co-administration of a vaccine in combination withadministration of a therapeutically effective amount of an IDO inhibitorof the present invention. In some embodiments, the vaccine is ananti-viral vaccine, including, for example, an anti-HIV vaccine. Inother embodiments, the vaccine is effective against tuberculosis ormalaria. In still other embodiments, the vaccine is a tumor vaccine(e.g., a vaccine effective against melanoma); the tumor vaccine maycomprise genetically modified tumor cells or a genetically modified cellline, including genetically modified tumor cells or a geneticallymodified cell line that has been transfected to expressgranulocyte-macrophage stimulating factor (GM-CSF). In particularembodiments, the vaccine includes one or more immunogenic peptidesand/or dendritic cells.

In some embodiments, the present invention contemplates methods of usingthe IDO inhibitors disclosed herein in combination with one or moreantimicrobial agents.

In certain embodiments drawn to treatment of an infection byadministering an IDO inhibitor and at least one additional therapeuticagent, a symptom of infection observed after administering both the IDOinhibitor and the additional therapeutic agent is improved over the samesymptom of infection observed after administering either alone. In someembodiments, the symptom of infection observed may be reduction in viralload, increase in CD4⁺ T cell count, decrease in opportunisticinfections, increased survival time, eradication of chronic infection,or a combination thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A, 1B, 1C and 1D provide structures and biological activity forcompounds described herein.

DETAILED DESCRIPTION OF THE INVENTION

Before the present invention is further described, it is to beunderstood that the invention is not limited to the particularembodiments set forth herein, and it is also to be understood that theterminology used herein is for the purpose of describing particularembodiments only, and is not intended to be limiting.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges, and are also encompassed within the invention, subjectto any specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention. Unlessdefined otherwise, all technical and scientific terms used herein havethe same meaning as commonly understood by one of ordinary skill in theart to which this invention belongs.

It must be noted that as used herein and in the appended claims, thesingular forms “a”, “an”, and “the” include plural referents unless thecontext clearly dictates otherwise. It is further noted that the claimsmay be drafted to exclude any optional element. As such, this statementis intended to serve as antecedent basis for use of such exclusiveterminology such as “solely”, “only” and the like in connection with therecitation of claim elements, or use of a “negative” limitation.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Further,the dates of publication provided may be different from the actualpublication dates, which may need to be independently confirmed.

General

Immune dysregulation is intimately associated with tumor evasion of thehost immune system, resulting in tumor growth and progression.Traditional treatment approaches comprising chemotherapy andradiotherapy are generally difficult for the patient to tolerate andbecome less effective as tumors evolve to survive such treatments. Byutilizing the patient's own immune system to identify and eliminatetumor cells, immunotherapy has the benefit of reduced toxicity. Asupregulation of the immunoregulatory enzyme indoleamine 2,3-dioxygenasecomprises one mechanism manipulated by tumors to promote growth, agents(e.g., small molecule compounds) that inhibit enzyme activity present apromising avenue for prophylaxis and/or treatment.

In addition, a large body of experimental data indicates a role for IDOinhibition in immunosuppression, tumor resistance and/or rejection,chronic infections, HIV-infection, and autoimmune diseases or disorders.Inhibition of IDO may also be an important treatment strategy forpatients with neurological or neuropsychiatric diseases or disorderssuch as depression. The compounds, compositions and methods hereinaddress the need for new classes of IDO modulators.

DEFINITIONS

Unless otherwise indicated, the following terms are intended to have themeaning set forth below. Other terms are defined elsewhere throughoutthe specification.

The term “alkyl”, by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, having the number of carbon atoms designated (i.e., C₁₋₈ meansone to eight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like.

The term “cycloalkyl” refers to hydrocarbon rings having the indicatednumber of ring atoms (e.g., C₃₋₆ cycloalkyl) and being fully saturatedor having no more than one double bond between ring vertices.“Cycloalkyl” is also meant to refer to bicyclic and polycyclichydrocarbon rings such as, for example, bicyclo[2.2.1]heptane,bicyclo[2.2.2]octane, etc.

The term “cycloheteroalkyl” refers to a cycloalkyl ring having theindicated number of ring vertices (or members) and having from one tofive heteroatoms selected from N, O, and S, which replace one to five ofthe carbon vertices, and wherein the nitrogen and sulfur atoms areoptionally oxidized, and the nitrogen atom(s) are optionallyquaternized. The cycloheteroalkyl may be a monocyclic, a bicyclic or apolycyclic ring system. Non limiting examples of cycloheteroalkyl groupsinclude pyrrolidine, imidazolidine, pyrazolidine, butyrolactam,valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide,piperidine, 1,4-dioxane, morpholine, thiomorpholine,thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran,pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran,tetrhydrothiophene, quinuclidine, and the like. A cycloheteroalkyl groupcan be attached to the remainder of the molecule through a ring carbonor a heteroatom.

As used herein, a wavy line, “

”, that intersects a single, double or triple bond in any chemicalstructure depicted herein, represent the point attachment of the single,double, or triple bond to the remainder of the molecule. Additionally, abond extending to the center of a ring (e.g., a phenyl ring) is meant toindicate attachment at any of the available ring vertices. One of skillin the art will understand that multiple substituents shown as beingattached to a ring will occupy ring vertices that provide stablecompounds and are otherwise sterically compatible. For a divalentcomponent, a representation is meant to include either orientation(forward or reverse). For example, the group “—C(O)NH—” is meant toinclude a linkage in either orientation: —C(O)NH— or —NHC(O)—, andsimilarly, “—O—CH₂CH₂—” is meant to include both —O—CH₂CH₂— and—CH₂CH₂—O—.

The terms “alkoxy”, “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. Additionally, for dialkylaminogroups, the alkyl portions can be the same or different and can also becombined to form a 3-7 membered ring with the nitrogen atom to whicheach is attached. Accordingly, a group represented as dialkylamino or—NR^(a)R^(b) is meant to include piperidinyl, pyrrolidinyl, morpholinyl,azetidinyl and the like.

The terms “halo” or “halogen”, by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl”, aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“C₁₋₄ haloalkyl” is mean to include trifluoromethyl,2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,typically aromatic, hydrocarbon group which can be a single ring ormultiple rings (up to three rings) which are fused together or linkedcovalently. Non-limiting examples of aryl groups include phenyl,naphthyl and biphenyl.

The term “heteroaryl” refers to aryl groups (or rings) that contain fromone to five heteroatoms selected from N, O, and S, wherein the nitrogenand sulfur atoms are optionally oxidized, and the nitrogen atom(s) areoptionally quaternized. A heteroaryl group can be attached to theremainder of the molecule through a heteroatom. Non-limiting examples ofheteroaryl groups include pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl,triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, cinnolinyl,phthalazinyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl,benzotriazolyl, benzisoxazolyl, isobenzofuryl, isoindolyl, indolizinyl,benzotriazinyl, thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl,imidazopyridines, benzothiaxolyl, benzofuranyl, benzothienyl, indolyl,quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl,imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl,pyrrolyl, thiazolyl, furyl, thienyl and the like. Substituents for aheteroaryl ring can be selected from the group of acceptablesubstituents described below.

The above terms (e.g., “alkyl”, “aryl” and “heteroaryl”), in someembodiments, will be optionally substituted. Selected substituents foreach type of radical are provided below.

Optional substituents for the alkyl radicals (including those groupsoften referred to as alkylene, alkenyl, alkynyl and cycloalkyl) can be avariety of groups selected from: halogen, —OR′, —NR′R″, —SR′,—SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR′S(O)₂R″, —CN and—NO₂ in a number ranging from zero to (2m′+1), where m′ is the totalnumber of carbon atoms in such radical. R′, R″ and R′″ eachindependently refer to hydrogen, unsubstituted C₁₋₈ alkyl, unsubstitutedaryl, aryl substituted with 1-3 halogens, unsubstituted C₁₋₈ alkyl, C₁₋₈alkoxy or C₁₋₈ thioalkoxy groups, or unsubstituted aryl-C₁₋₄ alkylgroups. When R′ and R″ are attached to the same nitrogen atom, they canbe combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or7-membered ring. For example, —NR′R″ is meant to include 1-pyrrolidinyland 4-morpholinyl.

Similarly, optional substituents for the aryl and heteroaryl groups arevaried and are generally selected from: -halogen, —OR′, —OC(O)R′,—NR′R″, —SR′, —R′, —CN, —NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″,—NR″C(O)R′, —NR″C(O)₂R′, —NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR′S(O)₂R″, —N₃,perfluoro(C₁-C₄)alkoxy, and perfluoro(C₁-C₄)alkyl, in a number rangingfrom zero to the total number of open valences on the aromatic ringsystem; and where R′, R″ and R′″ are independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl andC₂₋₈ alkynyl. Other suitable substituents include each of the above arylsubstituents attached to a ring atom by an alkylene tether of from 1-4carbon atoms.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula-A-(CH₂)_(r)—B—, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituent R′ in —NR′— and —S(O)₂NR′— is selected from hydrogen orunsubstituted C₁₋₆ alkyl.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si).

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of salts derived from pharmaceuticallyacceptable inorganic bases include aluminum, ammonium, calcium, copper,ferric, ferrous, lithium, magnesium, manganic, manganous, potassium,sodium, zinc and the like. Salts derived from pharmaceuticallyacceptable organic bases include salts of primary, secondary andtertiary amines, including substituted amines, cyclic amines,naturally-occurring amines and the like, such as arginine, betaine,caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, malonic, benzoic, succinic, suberic,fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, S. M. et al., “Pharmaceutical Salts”, J. Pharm. Sci., 66:1-19(1977)). Certain specific compounds of the present invention containboth basic and acidic functionalities that allow the compounds to beconverted into either base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers, regioisomers and individual isomers (e.g., separateenantiomers) are all intended to be encompassed within the scope of thepresent invention. When a stereochemical depiction is shown, it is meantto refer the compound in which one of the isomers is present andsubstantially free of the other isomer. “Substantially free of” anotherisomer indicates at least an 80/20 ratio of the two isomers, morepreferably 90/10, or 95/5 or more. In some embodiments, one of theisomers will be present in an amount of at least 99%.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. Unnatural proportions of an isotope may bedefined as ranging from the amount found in nature to an amountconsisting of 100% of the atom in question. For example, the compoundsmay incorporate radioactive isotopes, such as, for example, tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C), or non-radioactive isotopes,such as deuterium (²H) or carbon-13 (¹³C). Such isotopic variations canprovide additional utilities to those described elsewhere within thisapplication. For instance, isotopic variants of the compounds of theinvention may find additional utility, including but not limited to, asdiagnostic and/or imaging reagents, or as cytotoxic/radiotoxictherapeutic agents. Additionally, isotopic variants of the compounds ofthe invention can have altered pharmacokinetic and pharmacodynamiccharacteristics which can contribute to enhanced safety, tolerability orefficacy during treatment. All isotopic variations of the compounds ofthe present invention, whether radioactive or not, are intended to beencompassed within the scope of the present invention.

The terms “patient” or “subject” are used interchangeably to refer to ahuman or a non-human animal (e.g., a mammal).

The terms “administration”, “administer” and the like, as they apply to,for example, a subject, cell, tissue, organ, or biological fluid, referto contact of, for example, an inhibitor of IDO, a pharmaceuticalcomposition comprising same, or a diagnostic agent to the subject, cell,tissue, organ, or biological fluid. In the context of a cell,administration includes contact (e.g., in vitro or ex vivo) of a reagentto the cell, as well as contact of a reagent to a fluid, where the fluidis in contact with the cell.

The terms “treat”, “treating”, “treatment” and the like refer to acourse of action (such as administering an inhibitor of IDO or apharmaceutical composition comprising same) initiated after a disease,disorder or condition, or a symptom thereof, has been diagnosed,observed, and the like so as to eliminate, reduce, suppress, mitigate,or ameliorate, either temporarily or permanently, at least one of theunderlying causes of a disease, disorder, or condition afflicting asubject, or at least one of the symptoms associated with a disease,disorder, condition afflicting a subject. Thus, treatment includesinhibiting (e.g., arresting the development or further development ofthe disease, disorder or condition or clinical symptoms associationtherewith) an active disease.

The term “in need of treatment” as used herein refers to a judgment madeby a physician or other caregiver that a subject requires or willbenefit from treatment. This judgment is made based on a variety offactors that are in the realm of the physician's or caregiver'sexpertise.

The terms “prevent”, “preventing”, “prevention” and the like refer to acourse of action (such as administering an IDO inhibitor or apharmaceutical composition comprising same) initiated in a manner (e.g.,prior to the onset of a disease, disorder, condition or symptom thereof)so as to prevent, suppress, inhibit or reduce, either temporarily orpermanently, a subject's risk of developing a disease, disorder,condition or the like (as determined by, for example, the absence ofclinical symptoms) or delaying the onset thereof, generally in thecontext of a subject predisposed to having a particular disease,disorder or condition. In certain instances, the terms also refer toslowing the progression of the disease, disorder or condition orinhibiting progression thereof to a harmful or otherwise undesiredstate.

The term “in need of prevention” as used herein refers to a judgmentmade by a physician or other caregiver that a subject requires or willbenefit from preventative care. This judgment is made based on a varietyof factors that are in the realm of a physician's or caregiver'sexpertise.

The phrase “therapeutically effective amount” refers to theadministration of an agent to a subject, either alone or as part of apharmaceutical composition and either in a single dose or as part of aseries of doses, in an amount capable of having any detectable, positiveeffect on any symptom, aspect, or characteristic of a disease, disorderor condition when administered to the subject. The therapeuticallyeffective amount can be ascertained by measuring relevant physiologicaleffects, and it can be adjusted in connection with the dosing regimenand diagnostic analysis of the subject's condition, and the like. By wayof example, measurement of the serum level of and IDO inhibitor (or,e.g., a metabolite thereof) at a particular time post-administration maybe indicative of whether a therapeutically effective amount has beenused.

The phrase “in a sufficient amount to effect a change” means that thereis a detectable difference between a level of an indicator measuredbefore (e.g., a baseline level) and after administration of a particulartherapy. Indicators include any objective parameter (e.g., serumconcentration) or subjective parameter (e.g., a subject's feeling ofwell-being).

The term “small molecules” refers to chemical compounds having amolecular weight that is less than about 10 kDa, less than about 2 kDa,or less than about 1 kDa. Small molecules include, but are not limitedto, inorganic molecules, organic molecules, organic molecules containingan inorganic component, molecules comprising a radioactive atom, andsynthetic molecules. Therapeutically, a small molecule may be morepermeable to cells, less susceptible to degradation, and less likely toelicit an immune response than large molecules.

As used herein, the terms “IDO inhibitor”, “IDO blocker” and termssimilar thereto refer to agents capable of inhibiting the activity ofIDO, thereby reversing IDO-mediated immunosuppression. An IDO inhibitormay be a competitive, noncompetitive, or irreversible IDO inhibitor. “Acompetitive IDO inhibitor” is a compound that reversibly inhibits IDOenzyme activity at the catalytic site; “a noncompetitive IDO Inhibitor”is a compound that reversibly inhibits IDO enzyme activity at anon-catalytic site; and “an irreversible IDO inhibitor” is a compoundthat irreversibly eliminates IDO enzyme activity by forming a covalentbond (or other stable means of inhibiting enzyme function) with theenzyme. A number of IDO inhibitors are commercially available (e.g.,5-Br-4-Cl-indoxyl 1,3-diacetate and 1-methyl-DL-tryptophan (1 MT); bothavailable from Sigma-Aldrich, St. Louis, Mo.) and may be used as, forexample, “tool” or “reference” compounds

The term “ligand” refers to, for example, a peptide, a polypeptide, amembrane-associated or membrane-bound molecule, or a complex thereof,that can act as an agonist or antagonist of a receptor. A ligandencompasses natural and synthetic ligands, e.g., cytokines, cytokinevariants, analogs, muteins, and binding compositions derived fromantibodies, as well as small molecules. The term also encompasses anagent that is neither an agonist nor antagonist, but that can bind to areceptor without significantly influencing its biological properties,e.g., signaling or adhesion. Moreover, the term includes amembrane-bound ligand that has been changed by, e.g., chemical orrecombinant methods, to a soluble version of the membrane-bound ligand.A ligand or receptor may be entirely intracellular, that is, it mayreside in the cytosol, nucleus, or some other intracellular compartment.The complex of a ligand and receptor is termed a “ligand-receptorcomplex”.

The terms “inhibitors” and “antagonists”, or “activators” and “agonists”refer to inhibitory or activating molecules, respectively, for example,for the activation of, e.g., a ligand, receptor, cofactor, gene, cell,tissue, or organ. Inhibitors are molecules that decrease, block,prevent, delay activation, inactivate, desensitize, or down-regulate,e.g., a gene, protein, ligand, receptor, or cell. Activators aremolecules that increase, activate, facilitate, enhance activation,sensitize, or up-regulate, e.g., a gene, protein, ligand, receptor, orcell. An inhibitor may also be defined as a molecule that reduces,blocks, or inactivates a constitutive activity. An “agonist” is amolecule that interacts with a target to cause or promote an increase inthe activation of the target. An “antagonist” is a molecule that opposesthe action(s) of an agonist. An antagonist prevents, reduces, inhibits,or neutralizes the activity of an agonist, and an antagonist can alsoprevent, inhibit, or reduce constitutive activity of a target, e.g., atarget receptor, even where there is no identified agonist.

The terms “modulate”, “modulation” and the like refer to the ability ofa molecule (e.g., an activator or an inhibitor) to increase or decreasethe function or activity of IDO, either directly or indirectly. Amodulator may act alone, or it may use a cofactor, e.g., a protein,metal ion, or small molecule. Examples of modulators include smallmolecule compounds and other bioorganic molecules. Numerous libraries ofsmall molecule compounds (e.g., combinatorial libraries) arecommercially available and can serve as a starting point for identifyinga modulator. The skilled artisan is able to develop one or more assays(e.g., biochemical or cell-based assays) in which such compoundlibraries can be screened in order to identify one or more compoundshaving the desired properties; thereafter, the skilled medicinal chemistis able to optimize such one or more compounds by, for example,synthesizing and evaluating analogs and derivatives thereof. Syntheticand/or molecular modeling studies can also be utilized in theidentification of an Activator.

The “activity” of a molecule may describe or refer to the binding of themolecule to a ligand or to a receptor; to catalytic activity; to theability to stimulate gene expression or cell signaling, differentiation,or maturation; to antigenic activity; to the modulation of activities ofother molecules; and the like. The term “proliferative activity”encompasses an activity that promotes, that is necessary for, or that isspecifically associated with, for example, normal cell division, as wellas cancer, tumors, dysplasia, cell transformation, metastasis, andangiogenesis.

As used herein, “comparable”, “comparable activity”, “activitycomparable to”, “comparable effect”, “effect comparable to”, and thelike are relative terms that can be viewed quantitatively and/orqualitatively. The meaning of the terms is frequently dependent on thecontext in which they are used. By way of example, two agents that bothactivate a receptor can be viewed as having a comparable effect from aqualitative perspective, but the two agents can be viewed as lacking acomparable effect from a quantitative perspective if one agent is onlyable to achieve 20% of the activity of the other agent as determined inan art-accepted assay (e.g., a dose-response assay) or in anart-accepted animal model. When comparing one result to another result(e.g., one result to a reference standard), “comparable” frequently(though not always) means that one result deviates from a referencestandard by less than 35%, by less than 30%, by less than 25%, by lessthan 20%, by less than 15%, by less than 10%, by less than 7%, by lessthan 5%, by less than 4%, by less than 3%, by less than 2%, or by lessthan 1%. In particular embodiments, one result is comparable to areference standard if it deviates by less than 15%, by less than 10%, orby less than 5% from the reference standard. By way of example, but notlimitation, the activity or effect may refer to efficacy, stability,solubility, or immunogenicity.

“Substantially pure” indicates that a component makes up greater thanabout 50% of the total content of the composition, and typically greaterthan about 60% of the total polypeptide content. More typically,“substantially pure” refers to compositions in which at least 75%, atleast 85%, at least 90% or more of the total composition is thecomponent of interest. In some cases, the polypeptide will make upgreater than about 90%, or greater than about 95% of the total contentof the composition.

The terms “specifically binds” or “selectively binds”, when referring toa ligand/receptor, antibody/antigen, or other binding pair, indicates abinding reaction which is determinative of the presence of the proteinin a heterogeneous population of proteins and other biologics. Thus,under designated conditions, a specified ligand binds to a particularreceptor and does not bind in a significant amount to other proteinspresent in the sample. The antibody, or binding composition derived fromthe antigen-binding site of an antibody, of the contemplated methodbinds to its antigen, or a variant or mutein thereof, with an affinitythat is at least two-fold greater, at least ten times greater, at least20-times greater, or at least 100-times greater than the affinity withany other antibody, or binding composition derived therefrom. In aparticular embodiment, the antibody will have an affinity that isgreater than about 10⁹ liters/mol, as determined by, e.g., Scatchardanalysis (Munsen et al., Analyt. Biochem., 107:220-239 (1980)).

The term “response”, for example, of a cell, tissue, organ, or organism,encompasses a change in biochemical or physiological behavior, e.g.,concentration, density, adhesion, or migration within a biologicalcompartment, rate of gene expression, or state of differentiation, wherethe change is correlated with activation, stimulation, or treatment, orwith internal mechanisms such as genetic programming. In certaincontexts, the terms “activation”, “stimulation”, and the like refer tocell activation as regulated by internal mechanisms, as well as byexternal or environmental factors; whereas the terms “inhibition”,“down-regulation” and the like refer to the opposite effects.

The terms “polypeptide”, “peptide”, and “protein”, used interchangeablyherein, refer to a polymeric form of amino acids of any length, whichcan include genetically coded and non-genetically coded amino acids,chemically or biochemically modified or derivatized amino acids, andpolypeptides having modified polypeptide backbones. The terms includefusion proteins, including, but not limited to, fusion proteins with aheterologous amino acid sequence, fusion proteins with heterologous andhomologous leader sequences, with or without N-terminus methionineresidues; immunologically tagged proteins; and the like.

As used herein, the terms “variants” and “homologs” are usedinterchangeably to refer to amino acid or DNA sequences that are similarto reference amino acid or nucleic acid sequences, respectively. Theterm encompasses naturally-occurring variants andnon-naturally-occurring variants. Naturally-occurring variants includehomologs (polypeptides and nucleic acids that differ in amino acid ornucleotide sequence, respectively, from one species to another), andallelic variants (polypeptides and nucleic acids that differ in aminoacid or nucleotide sequence, respectively, from one individual toanother within a species). Thus, variants and homologs encompassnaturally occurring DNA sequences and proteins encoded thereby and theirisoforms, as well as splice variants of a protein or gene. The termsalso encompass nucleic acid sequences that vary in one or more basesfrom a naturally-occurring DNA sequence but still translate into anamino acid sequence that corresponds to the naturally-occurring proteindue to degeneracy of the genetic code. Non-naturally-occurring variantsand homologs include polypeptides and nucleic acids that comprise achange in amino acid or nucleotide sequence, respectively, where thechange in sequence is artificially introduced (e.g., muteins); forexample, the change is generated in the laboratory by human intervention(“hand of man”). Therefore, non-naturally occurring variants andhomologs may also refer to those that differ from thenaturally-occurring sequences by one or more conservative substitutionsand/or tags and/or conjugates.

The term “muteins” as used herein refers broadly to mutated recombinantproteins. These proteins usually carry single or multiple amino acidsubstitutions and are frequently derived from cloned genes that havebeen subjected to site-directed or random mutagenesis, or fromcompletely synthetic genes.

The terms “DNA”, “nucleic acid”, “nucleic acid molecule”,“polynucleotide” and the like are used interchangeably herein to referto a polymeric form of nucleotides of any length, eitherdeoxyribonucleotides or ribonucleotides, or analogs thereof.Non-limiting examples of polynucleotides include linear and circularnucleic acids, messenger RNA (mRNA), complementary DNA (cDNA),recombinant polynucleotides, vectors, probes, primers and the like.

Indoleamine 2,3-Dioxygenase

As previously alluded to, IDO is an immune regulatory enzyme that isnormally expressed in tumor cells and in activated immune cells. IDO isone of several immune response checkpoints that are involved in tumorimmune escape; thus, IDO inhibitors disrupt mechanisms by which tumorsevade the body's normal immune system.

IDO down-regulates the immune response mediated through oxidation oftryptophan. This results in inhibition of T-cell activation andinduction of T-cell apoptosis, creating an environment in whichtumor-specific cytotoxic T lymphocytes are rendered functionallyinactive or are no longer able to attack a subject's cancer cells.Therefore, therapeutic agents aimed at suppression of tryptophandegradation by inhibiting IDO activity are desirable. Inhibitors of IDOcan be used to activate T cells and therefore enhance T cell activationwhen the T cells are suppressed by pregnancy, malignancy or a virus suchas HIV Inhibition of IDO may also be an important treatment strategy forpatients with neurological or neuropsychiatric diseases or disorderssuch as depression. The compounds, compositions and methods herein helpmeet the current need for IDO modulators.

The expression of IDO is modulated by a complex array of signals, thusimplicating a number of different mechanisms of actions. For example,IDO may be induced by inhibition of DNA methyl transferases or histonedeacetylases. The NF-κB signaling pathway has also been implicated inIDO function. Inhibiting NF-κB activity blocks IDO expression andproduces robust anti-tumor responses that are both T cell- andIDO-dependent; alternatively, NF-κB activation (which may be effected byvarious factors such as interferon-γR1/-γR2 signaling andtoll-like-receptor activation) induces IDO gene expression.

Other mechanisms are involved with modulation of IDO function. By way ofexample, inhibitors of reactive oxidative species (ROS) may effectstabilization of IDO; IDO levels may be modulated by inhibition oractivation of pathways that are both downstream and upstream of IDO; andactivation of interferon-γ can activate an autocrine induction of IDO.

Studies indicate that the IDO pathway is active in many cancers, bothwithin tumor cells as a direct defense against T cell attack, and alsowithin antigen-presenting cells (APCs) in tumor-draining lymph nodesresulting in peripheral tolerance to tumor-associated antigens (TAAs).Cancers may use the IDO pathway to facilitate survival, growth,invasion, and metastasis of malignant cells expressing TAAs that mightotherwise be recognized and attacked by the immune system.

As alluded to herein, tryptophan catabolism in tumor tissue by therate-limiting enzyme IDO provides an opportunity for the use of IDOinhibitors as a therapeutic alternative to, or an additive with,conventional chemotherapy. However, certain cancers are capable ofcatabolizing tryptophan but are largely IDO-negative. Recent studiesindicate that the alternative enzymatic pathway of tryptophan catabolisminvolving tryptophan-2,3-dioxygenase (TDO) is also relevant in cancer.TDO, which is considered responsible for regulating systemic tryptophanlevels in the liver, is constitutively expressed in some cancers and isalso capable of suppressing antitumor immune responses (See, e.g.,Platten, M. et al., Cancer Res., 72(21):5435-5440 (Nov. 1, 2012)).

IDO is expressed in a wide variety of human tumors and tumor cell linesas well as in host APCs, which correlates with a worse clinicalprognosis. Therefore, inhibition of IDO may improve survival in cancerpatients with IDO-mediated immunosuppression. In comparison, TDO isexpressed in a wide variety of human tumors and tumor cell lines, andexpression of TDO is evident in advanced human glioblastomas. Theidentification of tumors expressing high levels of IDO or TDO may allowmore selective inhibition of the tryptophan-regulated immunosuppressivepathways. Alternatively, compounds inhibiting both IDO and TDO couldprovide the greatest coverage to prevent tumor escape by compensatoryexpression of the other tryptophan-degrading enzyme. Therefore, the useof dual IDO/TDO inhibitors or combinations of IDO- and TDO-specificinhibitors may prove to be a superior treatment alternative inimmunotherapy of cancer to block immunosuppression mediated bytryptophan metabolism.

Although a precise understanding of the underlying mechanism of actionby which the compounds of the present invention effect their activity isnot required to practice the invention, the compounds (or a subsetthereof) are believed to inhibit IDO function. Alternatively, thecompounds (or a subset thereof) may inhibit TDO function. The compounds(or a subset thereof) may also have inhibitory activity on both IDO andTDO function. Although the compounds of the invention are generallyreferred to herein as IDO inhibitors, it is to be understood that theterm “IDO inhibitors” encompasses compounds that act individuallythrough inhibition of TDO or IDO, and/or compounds that act throughinhibition of both IDO and TDO.

Identification of IDO Inhibitors Possessing Desirable Characteristics

The present invention is drawn, in part, to the identification ofinhibitors of IDO with at least one property or characteristic that isof therapeutic relevance. Candidate inhibitors may be identified byusing, for example, an art-accepted assay or model, examples of whichare described herein.

After identification, candidate inhibitors can be further evaluated byusing techniques that provide data regarding characteristics of theinhibitors (e.g., pharmacokinetic parameters, means of determiningsolubility or stability). Comparisons of the candidate inhibitors to areference standard (which may the “best-of-class” of current inhibitors)are indicative of the potential viability of such candidates.

Compounds of the Invention

As noted above, the present invention provides compounds represented byformula (I):

or a pharmaceutically acceptable salt, hydrate or solvate thereof,wherein,

-   the subscript n is 1 or 0;-   A is —C(O)—, —NH—, —SO₂—, —CH₂—, or —CHR³—;-   B is a bond, —C(O)—, —NH—, —CH₂—, or —CHR³—;-   T is a bond, —CH₂—, —NH—, —O—, —OCH₂—, —C(O)CH₂—, or —CR³R⁴—;    -   wherein when A is —NH— and B is —C(O)—, then T is other than        —C(R³)(R⁴)—;-   D is N or C(R⁵);-   E is N or C(R⁶);-   V is a bond, —O—, or —C(R^(5a))₂;-   G is an optionally substituted aryl, optionally substituted    heteroaryl, or an optionally substituted 9- or 10-membered fused    bicyclic heteroaryl;-   J¹ is CH, N or C(R²), when R² is attached to the ring vertex    identified as J¹;-   R¹ and R² are independently hydrogen, halogen, optionally    substituted C₁-C₄ haloalkyl, optionally substituted C₃-C₆    cycloalkyl, optionally substituted 3- to 6-membered    cycloheteroalkyl, optionally substituted phenyl, optionally    substituted heteroaryl, optionally substituted C₁-C₄ alkyl,    optionally substituted C₁-C₄ alkoxy, CN, SO₂NH₂, NHSO₂CH₃, NHSO₂CF₃,    OCF₃, SO₂CH₃, SO₂CF₃, or CONH₂, and when R¹ and R² are on adjacent    vertices of a phenyl ring they may be joined together to form a 5-    or 6-membered cycloheteroalkyl ring having one or two ring vertices    independently selected from O, N and S, wherein said    cycloheteroalkyl ring is optionally substituted with from one to    three members selected from fluoro and C₁-C₃ alkyl;-   R³ and R⁴ are independently hydrogen, optionally substituted C₁-C₆    alkyl, optionally substituted C₁-C₆ haloalkyl, fluorine, OH, CN,    CO₂H, C(O)NH₂, N(R^(5a))₂, optionally substituted —O—C₁-C₆ alkyl,    —(CR⁵R⁵)_(m)—OH, —(CR⁵R⁵)_(m)—CO₂H, —(CR⁵R⁵)_(m)—C(O)NH₂,    —(CR⁵R⁵)_(m)—C(O)NHR^(5a), —(CR⁵R⁵)_(m)N(R^(5a))₂,    —NH(CR⁵R⁵)_(m)CO₂H or —NH(CR⁵R⁵)_(m)—C(O)NH₂;-   each R⁵ is independently H, F, OH, optionally substituted C₁-C₆    alkyl or optionally substituted —O—C₁-C₆ alkyl;-   each R^(5a) is independently H, or optionally substituted C₁-C₆    alkyl;-   R⁶ is H, OH, F, optionally substituted C₁-C₆ alkyl, optionally    substituted —O—C₁-C₆ alkyl, or —N(R^(5a))₂;-   and each m is independently 1, 2, or 3.

In some embodiments, the compounds provided herein have the formula(Ia):

In some selected embodiments of formula (Ia), compounds are providedhaving formulae (Ia1), (Ia2) or (Ia3):

In some embodiments, the compounds provided herein have the formula(Ib), (Ic), or (Id):

In some embodiments, the compounds provided herein have the formula(Ie):

In some selected embodiments of formula (Ie), compounds are providedhaving formulae (Ie1):

In some embodiments, the compounds provided herein have the formula(If), (Ig), or (Ih):

In some embodiments, the compounds provided herein have the formula (Ii)or (Ij):

For each of the above formulae (Ia), (Ia1), (Ia2), (Ia3), (Ib), (Ic),(Id), (Ie), (Ie1), (If), (Ig), (Ih), (Ii) and (Ij), each of thesubscript, letters, J¹, R¹, R², R³, R⁴ and R⁵ have the meanings providedwith reference to formula (I), unless noted otherwise.

In one group of selected embodiments, any one compound of FIG. 1 isprovided.

In another group of selected embodiments, any one compound of FIG. 1 isprovided having an activity level identified as “A” or “B”.

In another group of selected embodiments, any one compound of FIG. 1 isprovided having an activity level identified as “A”.

Methods of Synthesis

The compounds of the present invention may be prepared from startingmaterials which are known in the chemical literature or are commerciallyavailable by methods such as those illustrated in the following Schemesutilizing chemical transformations known to those skilled in the art oforganic chemistry. Solvents, temperatures, pressures, and other reactionconditions may readily be selected by one of ordinary skill in the art.These Schemes are illustrative and are not meant to limit the possibletechniques one skilled in the art may use to manufacture compoundsdisclosed herein. Different methods may be evident to those skilled inthe art. Additionally, the various steps in the synthesis may beperformed in an alternate sequence or order to give the desiredcompound(s). Further, the representation of the reactions in theseSchemes as discrete steps does not preclude their being performed intandem, either by telescoping multiple steps in the same reaction vesselor by performing multiple steps without purifying or characterizing theintermediate(s). In addition, many of the compounds prepared by themethods below can be further modified using conventional chemistry wellknown to those skilled in the art. All documents cited herein areincorporated herein by reference in their entirety.

References to many of these chemical transformations employed herein canbe found in Smith, M. B. et al., March's Advanced Organic ChemistryReactions, Mechanisms, and Structure, Fifth Edition, Wiley-Interscience,New York (2001), or other standard texts on the topic of syntheticorganic chemistry. Certain transformations may require that reactivefunctional groups be masked by protecting group(s). A convenientreference which provides conditions for introduction, removal, andrelative susceptibility to reaction conditions of these groups isGreene, T. W. et al., Protective Groups in Organic Synthesis, ThirdEdition, Wiley-Interscience, New York (1999).

Scheme 1 Reverse Amides and Sulfonamides, Cycloalkyl Core, Direct orO-Linked G

Treatment of a phosphonoacetate ester (III), with a base such as sodiumhydride in a solvent such as THF (Scheme 1) followed by a ketone of thegeneral structure II affords a trisubstituted olefin. Substitutedanalogs of III (R³ is not H) afford tetrasubstituted olefins. Thismethod and additional methods described below are transformationsfamiliar to those skilled in the art of organic/medicinal chemistry.Alternative methods for olefination and the transformations describedbelow are known and will be selected by one skilled in the art based ontheir applicability to the specific substrate under consideration.Reduction is accomplished by stirring or shaking a solution of theolefin in a suitable solvent under an atmosphere or more of H₂ in thepresence of a catalyst, normally palladium on carbon. Hydrolysis of theketal group affords a ketone of the general structure IV. Typically,this is accomplished by heating with an aqueous acid such as HCl in thepresence of a co-solvent such as THF. In addition to the cyclic ethyleneglycol-based ketal shown, other cyclic and acyclic ketal protectinggroups could be used. Ketones are deprotonated with bases such as LiHMDSand react with N-phenyltrifluoromethanesulfonimide or similar reagentsto afford triflates of the general structure V. These triflatesparticipate in Suzuki couplings (T. Ishiyama, M. Murata, N. Miyaura, J.Org. Chem., 1995, 60, 7508-7510) with boronic acids or esters G-B(OR)₂to afford coupled products. Many variations on this reaction are known,but generally it involves heating the two substrates and a catalyst suchas (Ph₃P)₄Pd in a solvent such as DMF with a base such as aq. potassiumcarbonate. Reduction of the olefin provides intermediate VI (where V=abond and E=CH). Ketones IV can be reduced by NaBH₄ or similar hydridereducing agents to afford alcohols of the general structure VII. Thesealcohols can be deprotonated with bases such as sodium hydride or KHMDS,and the resulting alkoxides can react with aryl or heteroaryl halidesunder SNAr conditions to afford additional intermediates VI (where V=Oand E=CH). Alternatively, reaction can be accomplished by activation ofalcohol VII with DIAD, DEAD, or a related azodicarboxylate and atrialkyl or triarylphosphine and coupling with a phenol or relatedheteroaryl GOH. (Mitsunobu reaction) Intermediates VI (and laterintermediates) may be obtained as mixtures of cis and trans isomers.Methods for control of the stereochemical outcome of the above reactionsare known to those familiar in the art of organic/medicinal chemistry.Additionally, methods for the separation of these isomers are known anddescribed in detail in the synthetic examples. If required, the group R⁴can be appended by alkylation of intermediate VI. Methods for control ofthe absolute stereochemistry of the resulting asymmetric center areknown to those familiar with the art, as are chiral separation methods.Saponification of the ester by heating with aq. LiOH or a similar base,generally in the presence of an organic co-solvent such as THF affordscarboxylic acids VIII. Acids VIII can be rearranged, usually by heatingwith DPPA and triethylamine (Curtius and related rearrangements), andthe intermediate isocyanates react with aq. base to afford primaryamines IX. These can react with electrophiles including, but not limitedto acid chlorides and sulfonyl chlorides XVIa to afford compounds of theinvention I. Another means of preparing compounds I from IX where A isCO uses the carboxylic acid derivative of XVIa and peptide couplingreagents. For a recent review of peptide coupling methods see: AymanEl-Faham and Fernando Albericio. Chem. Rev. 2011, 111, 6557-6602.

Scheme 2 Reverse Amides and Sulfonamides, Piperidine Core, Direct orC-Linked G

Piperidine and pyrrolidine esters X are known compounds and can undergoS_(N)Ar (V=a bond) and N-alklyation (V=C(R^(5a))₂) reactions to affordintermediates VI (E=N). These intermediates may be transformed tocompounds of the invention I as shown in Scheme I.

Scheme 3 Reverse Amides and Sulfonamides, Cycloalkyl or Piperidine Core,Direct C, or O-Linked G, Alternate Method

Scheme 3 illustrates a method of making intermediate VI by performingthe steps of Scheme 1 in a different order. Intermediate II can beconverted to a triflate as described above and coupled with a boronicacid or ester G-B(OR)₂ to give intermediate XI (V=a bond). Analogs whereV=O can be prepared by reduction of ketone II and conversion to theether XI as described above. Ketal hydrolysis affords ketone XII.Transformation to intermediate VI is accomplished by olefination andreduction as described above. Intermediates XII in which E=N can beprepared from ketones XIII using SNAr (V=a bond) or alkylation(V=C(R^(5a))₂) chemistry.

Scheme 4 Truncated Normal Amides, Cycloalkyl or Piperidine Core, Direct,C, or O-Linked G

Scheme 4 illustrates a method of making additional compounds of theinvention. Ketone XII can be transformed into a nitrile by the use ofTosMIC (Van Leusen reaction). Hydrolysis of the nitrile affords acid XIVwhich can be converted into compounds of the invention I by treatmentwith amines under amide coupling conditions.

Scheme 5 Ureas and Phenylaceticamides of Cyclohexylamine or4-aminopiperidine, Direct, C, or O-Linked G

Scheme 5 illustrates a method of making additional compounds of theinvention. Ketones XII can be transformed into primary amines XV byreductive amination with ammonia or an ammonium salt. This reaction isgenerally performed using sodium cyanoborohydride in an alcoholicsolvent. Treatment with isocyanates XVIc affords ureas which arecompounds of the invention I. Arylacetamides which are compounds of theinvention I are obtained by coupling amine XV with arylacetic acids XVIdusing a reagent such as Bop and a tertiary amine base in a solvent suchas DMF.

Scheme 6 Control of Absolute Stereochemistry in the Conversion of VI toVIII

Scheme 6 illustrates a method for controlling the absolutestereochemistry of intermediate VIII and materials arising from it.Saponification of esters VI provides carboxylic acids XVII. Treatment ofthese acids with an acid chloride such as pivaloyl chloride provides amixed anhydride intermediate. In a separate vessel, an optically pureoxazolidinone of known stereochemistry and general structure XVIII isdeprotonated by treatment with a strong base such as n-BuLi. Theseactivated species are combined to form the acyloxazolidinone XIX whichis deprotonated by bases such as NaHMDS. Alkylation of the resultingenolate proceeds with predictable control of stereochemistry at thenewly-formed center to provide materials XX. Removal of the chiralauxiliary to give optically-active carboxylic acids VIII is accomplishedby treatment with a solution of basic hydrogen peroxide. For a review ofthe history and scope scope of this reaction see: D. A. Evans, M. D.Ennis, D. J. Mathre. J. Am. Chem. Soc., 1982, 104 (6), pp 1737-1739.

Scheme 7 Synthesis of Compounds of the Invention (I) where R⁶=OH

As shown in Scheme 7, a ketone of general structure IV can be treatedwith a the lithiate G-Li, which can be generated by several methodswell-known to one skilled in the art, to produce a tertiary alcohol ofgeneral structure XXI. The ester of general structure XXXI can beconverted to a compound of general structure I via methods alreadydescribed herein. Alternatively, the ketone IV can be treated with theorganometallic MgBr—V-G (Grignard reagent) to give a tertiary alcohol ofgeneral structure XXII.

Scheme 8 Synthesis of Compounds of the Invention (I) where R⁵=OH

As shown in Scheme 8, a ketone of general structure XII can be treatedwith a halo acetate where X=Br in the presence of Zinc metal(Reformatsky reaction) to give the a tertiary alcohol of generalstructure XXIII. The ester XXIII can be converted to a compound of theInvention (I) by methods already described herein.

In addition to the above general schemes, the compounds described hereincan be prepared by representative methods as provided in the Examplesbelow.

Modifications to Enhance Inhibitor Characteristics

It is frequently beneficial, and sometimes imperative, to improve one ofmore physical properties of the treatment modalities disclosed hereinand/or the manner in which they are administered. Improvements ofphysical properties include, for example, methods of increasing watersolubility, bioavailability, serum half-life, and/or therapeutichalf-life; and/or modulating biological activity.

Modifications known in the art include pegylation, Fc-fusion and albuminfusion. Although generally associated with large molecule agents (e.g.,polypeptides), such modifications have recently been evaluated withparticular small molecules. By way of example, Chiang, M. et al., (J.Am. Chem. Soc., 136(9):3370-3373 (2014)) describe a small moleculeagonist of the adenosine 2a receptor conjugated to the immunoglobulin Fedomain. The small molecule-Fc conjugate retained potent Fc receptor andadenosine 2a receptor interactions and showed superior propertiescompared to the unconjugated small molecule. Covalent attachment of PEGmolecules to small molecule therapeutics has also been described (Li, W.et al., Progress in Polymer Science, 38:421-444 (2013)).

Therapeutic and Prophylactic Uses

The present invention contemplates the use of the TDO inhibitorsdescribed herein in the treatment or prevention of a broad range ofdiseases, disorders and/or conditions, and/or the symptoms thereof.While particular uses are described in detail hereafter, it is to beunderstood that the present invention is not so limited. Furthermore,although general categories of particular diseases, disorders andconditions are set forth hereafter, some of the diseases, disorders andconditions may be a member of more than one category, and others may notbe a member of any of the disclosed categories.

Oncology-Related Disorders.

In accordance with the present invention, an IDO inhibitor can be usedto treat or prevent a proliferative condition or disorder, including acancer, for example, cancer of the uterus, cervix, breast, prostate,testes, gastrointestinal tract (e.g., esophagus, oropharynx, stomach,small or large intestines, colon, or rectum), kidney, renal cell,bladder, bone, bone marrow, skin, head or neck, liver, gall bladder,heart, lung, pancreas, salivary gland, adrenal gland, thyroid, brain(e.g., gliomas), ganglia, central nervous system (CNS) and peripheralnervous system (PNS), and cancers of the hematopoietic system and theimmune system (e.g., spleen or thymus). The present invention alsoprovides methods of treating or preventing other cancer-relateddiseases, disorders or conditions, including, for example, immunogenictumors, non-immunogenic tumors, dormant tumors, virus-induced cancers(e.g., epithelial cell cancers, endothelial cell cancers, squamous cellcarcinomas and papillomavirus), adenocarcinomas, lymphomas, carcinomas,melanomas, leukemias, myelomas, sarcomas, teratocarcinomas,chemically-induced cancers, metastasis, and angiogenesis. The inventioncontemplates reducing tolerance to a tumor cell or cancer cell antigen,e.g., by modulating activity of a regulatory T-cell and/or a CD8+ T-cell(see, e.g., Ramirez-Montagut et al., Oncogene, 22:3180-3187 (2003); andSawaya et al., New Engl. J. Med., 349:1501-1509 (2003)). In particularembodiments, the tumor or cancer is colon cancer, ovarian cancer, breastcancer, melanoma, lung cancer, glioblastoma, or leukemia. The use of theterm(s) cancer-related diseases, disorders and conditions is meant torefer broadly to conditions that are associated, directly or indirectly,with cancer, and includes, e.g., angiogenesis and precancerousconditions such as dysplasia.

In some embodiments, the present invention provides methods for treatinga proliferative condition, cancer, tumor, or precancerous condition withan IDO inhibitor and at least one additional therapeutic or diagnosticagent, examples of which are set forth elsewhere herein.

Immune- and Inflammatory-Related Disorders.

As used herein, terms such as “immune disease”, “immune condition”,“immune disorder”, “inflammatory disease”, “inflammatory condition”,“inflammatory disorder” and the like are meant to broadly encompass anyimmune- or inflammatory-related condition (e.g., pathologicalinflammation and autoimmune diseases). Such conditions frequently areinextricably intertwined with other diseases, disorders and conditions.By way of example, an “immune condition” may refer to proliferativeconditions, such as cancer, tumors, and angiogenesis; includinginfections (acute and chronic), tumors, and cancers that resisteradication by the immune system.

A non-limiting list of immune- and inflammatory-related diseases,disorders and conditions which may be treated or prevented with thecompounds and compositions of the present invention include, arthritis(e.g., rheumatoid arthritis), kidney failure, lupus, asthma, psoriasis,colitis, pancreatitis, allergies, fibrosis, surgical complications(e.g., where inflammatory cytokines prevent healing), anemia, andfibromyalgia. Other diseases and disorders which may be associated withchronic inflammation include Alzheimer's disease, congestive heartfailure, stroke, aortic valve stenosis, arteriosclerosis, osteoporosis,Parkinson's disease, infections, inflammatory bowel disease (e.g.,Crohn's disease and ulcerative colitis), allergic contact dermatitis andother eczemas, systemic sclerosis, transplantation and multiplesclerosis.

Among other immune-related disorders, it is contemplated that inhibitionof IDO function may also play a role in immunologic tolerance andprevention of fetal rejection in utero.

In some embodiments, an IDO inhibitor described herein can be combinedwith an immunosuppressive agent to reduce the number of immune effectorcells.

Some of the aforementioned diseases, disorders and conditions for whichan IDO inhibitor may be particularly efficacious (due to, for example,limitations of current therapies) are described in more detailhereafter.

Rheumatoid Arthritis (RA), which is generally characterized by chronicinflammation in the membrane lining (the synovium) of the joints,affects approximately 1% of the U.S. population (˜2.1 million people).Further understanding of the role of cytokines, including TNF-α andIL-1, in the inflammatory process has enabled the development andintroduction of a new class of disease-modifying antirheumatic drugs(DMARDs). Agents (some of which overlap with treatment modalities forRA) include ENBREL® (etanercept), REMICADE® (infliximab), HUMIRA®(adalimumab) and KINERET® (anakinra). Though some of these agentsrelieve symptoms, inhibit progression of structural damage, and improvephysical function in particular patient populations, there is still aneed for alternative agents with improved efficacy, complementarymechanisms of action, and fewer/less severe adverse effects.

Psoriasis, a constellation of common immune-mediated chronic skindiseases, affects more than 4.5 million people in the U.S., of which 1.5million are considered to have a moderate-to severe form of the disease.Moreover, over 10% of patients with psoriasis develop psoriaticarthritis, which damages the bone and connective tissue around thejoints. An improved understanding of the underlying physiology ofpsoriasis has resulted in the introduction of agents that, for example,target the activity of T lymphocytes and cytokines responsible for theinflammatory nature of the disease. Such agents include the TNF-αinhibitors (also used in the treatment of rheumatoid arthritis (RA)),including ENBREL® (etanercept), REMICADE® (infliximab) and HUMIRA®(adalimumab)), and T-cell inhibitors such as AMEVIVE® (alefacept) andRAPTIVA® (efalizumab). Though several of these agents are effective tosome extent in certain patient populations, none have been shown toeffectively treat all patients.

Subjects suffering from multiple sclerosis (MS), a seriouslydebilitating autoimmune disease comprising multiple areas ofinflammation and scarring of the myelin in the brain and spinal cord,may be particularly helped by the IDO inhibitors described herein, ascurrent treatments only alleviate symptoms or delay the progression ofdisability.

Similarly, the IDO inhibitors may be particularly advantageous forsubjects afflicted with neurodegenerative disorders, such as Alzheimer'sdisease (AD), a brain disorder that seriously impairs patients' thought,memory, and language processes; and Parkinson's disease (PD), aprogressive disorder of the CNS characterized by, for example, abnormalmovement, rigidity and tremor. These disorders are progressive anddebilitating, and no curative agents are available.

Viral-Related Disorders.

The present invention contemplates the use of the IDO inhibitors in thetreatment and/or prevention of any viral disease, disorder or conditionfor which treatment with an IDO inhibitor may be beneficial. Inparticular embodiments, the viral disorder is a chronic viral disorder.Examples of viral diseases, disorders and conditions that arecontemplated include, but are not limited to, hepatitis B virus (HBV),hepatitis C virus (HCV), human papilloma virus (HPV), HIV, AIDS(including its manifestations such as cachexia, dementia, and diarrhea),herpes simplex virus (HSV), Epstein-Barr virus (EBV), varicella zostervirus, coxsackie virus, and cytomegalovirus (CMV).

Bacterial- and Parasitic-Related Disorders.

Embodiments of the present invention contemplate the administration ofthe IDO inhibitors described herein to a subject for the treatment of abacterial infection, for example, a Mycobacterium infection (e.g.,Mycobacterium leprae or Mycobacterium tuberculosis) or an infectioncaused by Listeria monocytogenes or Toxplasma gondii. Other embodimentscontemplate the treatment of a parasitic infection including, but notlimited to, Leishmania donovani, Leishmania tropica, Leishmania major,Leishmania aethiopica, Leishmania mexicana, Plasmodium falciparum,Plasmodium vivax, Plasmodium ovale, or Plasmodium malariae. Frequently,anti-parasitic therapy is administered prophylactically (e.g., before asubject travels to an area with a high frequency of parasiticinfection).

Pharmaceutical Compositions

The IDO inhibitors of the present invention may be in the form ofcompositions suitable for administration to a subject. In general, suchcompositions are “pharmaceutical compositions” comprising an IDOinhibitor(s) and one or more pharmaceutically acceptable orphysiologically acceptable diluents, carriers or excipients. In certainembodiments, the IDO inhibitors are present in a therapeuticallyacceptable amount. The pharmaceutical compositions may be used in themethods of the present invention; thus, for example, the pharmaceuticalcompositions can be administered ex vivo or in vivo to a subject inorder to practice the therapeutic and prophylactic methods and usesdescribed herein.

The pharmaceutical compositions of the present invention can beformulated to be compatible with the intended method or route ofadministration; exemplary routes of administration are set forth herein.Furthermore, the pharmaceutical compositions may be used in combinationwith other therapeutically active agents or compounds as describedherein in order to treat or prevent the diseases, disorders andconditions as contemplated by the present invention.

The pharmaceutical compositions containing the active ingredient (e.g.,an inhibitor of IDO function) may be in a form suitable for oral use,for example, as tablets, capsules, troches, lozenges, aqueous or oilysuspensions, dispersible powders or granules, emulsions, hard or softcapsules, or syrups, solutions, microbeads or elixirs. Pharmaceuticalcompositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions, and such compositions may contain one or more agents suchas, for example, sweetening agents, flavoring agents, coloring agentsand preserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets, capsules and the like contain theactive ingredient in admixture with non-toxic pharmaceuticallyacceptable excipients which are suitable for the manufacture of tablets.These excipients may be, for example, diluents, such as calciumcarbonate, sodium carbonate, lactose, calcium phosphate or sodiumphosphate; granulating and disintegrating agents, for example, cornstarch, or alginic acid; binding agents, for example, starch, gelatin oracacia, and lubricating agents, for example, magnesium stearate, stearicacid or talc.

The tablets, capsules and the like suitable for oral administration maybe uncoated or coated by known techniques to delay disintegration andabsorption in the gastrointestinal tract and thereby provide a sustainedaction. For example, a time-delay material such as glyceryl monostearateor glyceryl distearate may be employed. They may also be coated bytechniques known in the art to form osmotic therapeutic tablets forcontrolled release. Additional agents include biodegradable orbiocompatible particles or a polymeric substance such as polyesters,polyamine acids, hydrogel, polyvinyl pyrrolidone, polyanhydrides,polyglycolic acid, ethylene-vinylacetate, methylcellulose,carboxymethylcellulose, protamine sulfate, or lactide/glycolidecopolymers, polylactide/glycolide copolymers, or ethylenevinylacetatecopolymers in order to control delivery of an administered composition.For example, the oral agent can be entrapped in microcapsules preparedby coacervation techniques or by interfacial polymerization, by the useof hydroxymethylcellulose or gelatin-microcapsules orpoly(methylmethacrolate) microcapsules, respectively, or in a colloiddrug delivery system. Colloidal dispersion systems include macromoleculecomplexes, nano-capsules, microspheres, microbeads, and lipid-basedsystems, including oil-in-water emulsions, micelles, mixed micelles, andliposomes. Methods for the preparation of the above-mentionedformulations will be apparent to those skilled in the art.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate, kaolin ormicrocrystalline cellulose, or as soft gelatin capsules wherein theactive ingredient is mixed with water or an oil medium, for example,peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture thereof. Such excipients can besuspending agents, for example, sodium carboxymethylcellulose,methylcellulose, hydroxy-propylmethylcellulose, sodium alginate,polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing orwetting agents, for example, a naturally-occurring phosphatide (e.g.,lecithin), or condensation products of an alkylene oxide with fattyacids (e.g., polyoxyethylene stearate), or condensation products ofethylene oxide with long chain aliphatic alcohols (e.g., forheptadecaethyleneoxycetanol), or condensation products of ethylene oxidewith partial esters derived from fatty acids and a hexitol (e.g.,polyoxyethylene sorbitol monooleate), or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides (e.g., polyethylene sorbitan monooleate). The aqueoussuspensions may also contain one or more preservatives.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example, arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example, beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified herein.

The pharmaceutical compositions of the present invention may also be inthe form of oil-in-water emulsions. The oily phase may be a vegetableoil, for example, olive oil or arachis oil, or a mineral oil, forexample, liquid paraffin, or mixtures of these. Suitable emulsifyingagents may be naturally occurring gums, for example, gum acacia or gumtragacanth; naturally occurring phosphatides, for example, soy bean,lecithin, and esters or partial esters derived from fatty acids; hexitolanhydrides, for example, sorbitan monooleate; and condensation productsof partial esters with ethylene oxide, for example, polyoxyethylenesorbitan monooleate.

Formulations can also include carriers to protect the compositionagainst rapid degradation or elimination from the body, such as acontrolled release formulation, including implants, liposomes,hydrogels, prodrugs and microencapsulated delivery systems. For example,a time delay material such as glyceryl monostearate or glyceryl stearatealone, or in combination with a wax, may be employed.

The pharmaceutical compositions typically comprise a therapeuticallyeffective amount of an IDO inhibitor contemplated by the presentinvention and one or more pharmaceutically and physiologicallyacceptable formulation agents. Suitable pharmaceutically acceptable orphysiologically acceptable diluents, carriers or excipients include, butare not limited to, antioxidants (e.g., ascorbic acid and sodiumbisulfate), preservatives (e.g., benzyl alcohol, methyl parabens, ethylor n-propyl, p-hydroxybenzoate), emulsifying agents, suspending agents,dispersing agents, solvents, fillers, bulking agents, detergents,buffers, vehicles, diluents, and/or adjuvants. For example, a suitablevehicle may be physiological saline solution or citrate buffered saline,possibly supplemented with other materials common in pharmaceuticalcompositions for parenteral administration. Neutral buffered saline orsaline mixed with serum albumin are further exemplary vehicles. Thoseskilled in the art will readily recognize a variety of buffers that canbe used in the pharmaceutical compositions and dosage forms contemplatedherein. Typical buffers include, but are not limited to,pharmaceutically acceptable weak acids, weak bases, or mixtures thereof.As an example, the buffer components can be water soluble materials suchas phosphoric acid, tartaric acids, lactic acid, succinic acid, citricacid, acetic acid, ascorbic acid, aspartic acid, glutamic acid, andsalts thereof. Acceptable buffering agents include, for example, a Trisbuffer, N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid) (HEPES),2-(N-morpholino)ethanesulfonic acid (MES),2-(N-morpholino)ethanesulfonic acid sodium salt (MES),3-(N-morpholino)propanesulfonic acid (MOPS), andN-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid (TAPS).

After a pharmaceutical composition has been formulated, it may be storedin sterile vials as a solution, suspension, gel, emulsion, solid, ordehydrated or lyophilized powder. Such formulations may be stored eitherin a ready-to-use form, a lyophilized form requiring reconstitutionprior to use, a liquid form requiring dilution prior to use, or otheracceptable form. In some embodiments, the pharmaceutical composition isprovided in a single-use container (e.g., a single-use vial, ampoule,syringe, or autoinjector (similar to, e.g., an EPIPEN®)), whereas amulti-use container (e.g., a multi-use vial) is provided in otherembodiments. Any drug delivery apparatus may be used to deliver and IDOinhibitor, including implants (e.g., implantable pumps) and cathetersystems, slow injection pumps and devices, all of which are well knownto the skilled artisan. Depot injections, which are generallyadministered subcutaneously or intramuscularly, may also be utilized torelease the polypeptides disclosed herein over a defined period of time.Depot injections are usually either solid- or oil-based and generallycomprise at least one of the formulation components set forth herein.One of ordinary skill in the art is familiar with possible formulationsand uses of depot injections.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents mentioned herein. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example, as a solution in 1,3-butane diol. Acceptable diluents,solvents and dispersion media that may be employed include water,Ringer's solution, isotonic sodium chloride solution, CREMOPHOR® EL(BASF, Parsippany, N.J.) or phosphate buffered saline (PBS), ethanol,polyol (e.g., glycerol, propylene glycol, and liquid polyethyleneglycol), and suitable mixtures thereof. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose any bland fixed oil may be employed, including synthetic mono-or diglycerides. Moreover, fatty acids such as oleic acid, find use inthe preparation of injectables. Prolonged absorption of particularinjectable formulations can be achieved by including an agent thatdelays absorption (e.g., aluminum monostearate or gelatin).

The present invention contemplates the administration of the IDOinhibitors in the form of suppositories for rectal administration. Thesuppositories can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials include, but are not limited to,cocoa butter and polyethylene glycols.

The IDO inhibitors contemplated by the present invention may be in theform of any other suitable pharmaceutical composition (e.g., sprays fornasal or inhalation use) currently known or developed in the future.

The concentration of a polypeptide or fragment thereof in a formulationcan vary widely (e.g., from less than about 0.1%, usually at or at leastabout 2% to as much as 20% to 50% or more by weight) and will usually beselected primarily based on fluid volumes, viscosities, andsubject-based factors in accordance with, for example, the particularmode of administration selected.

Routes of Administration

The present invention contemplates the administration of IDO inhibitors,and compositions thereof, in any appropriate manner. Suitable routes ofadministration include oral, parenteral (e.g., intramuscular,intravenous, subcutaneous (e.g., injection or implant), intraperitoneal,intracisternal, intraarticular, intraperitoneal, intracerebral(intraparenchymal) and intracerebroventricular), nasal, vaginal,sublingual, intraocular, rectal, topical (e.g., transdermal), sublingualand inhalation. Depot injections, which are generally administeredsubcutaneously or intramuscularly, may also be utilized to release theIDO inhibitors disclosed herein over a defined period of time.

Particular embodiments of the present invention contemplate oraladministration.

Combination Therapy

The present invention contemplates the use of IDO inhibitors incombination with one or more active therapeutic agents (e.g.,chemotherapeutic agents) or other prophylactic or therapeutic modalities(e.g., radiation). In such combination therapy, the various activeagents frequently have different, complementary mechanisms of action.Such combination therapy may be especially advantageous by allowing adose reduction of one or more of the agents, thereby reducing oreliminating the adverse effects associated with one or more of theagents. Furthermore, such combination therapy may have a synergistictherapeutic or prophylactic effect on the underlying disease, disorder,or condition.

As used herein, “combination” is meant to include therapies that can beadministered separately, for example, formulated separately for separateadministration (e.g., as may be provided in a kit), and therapies thatcan be administered together in a single formulation (i.e., a“co-formulation”).

In certain embodiments, the IDO inhibitors are administered or appliedsequentially, e.g., where one agent is administered prior to one or moreother agents. In other embodiments, the IDO inhibitors are administeredsimultaneously, e.g., where two or more agents are administered at orabout the same time; the two or more agents may be present in two ormore separate formulations or combined into a single formulation (i.e.,a co-formulation). Regardless of whether the two or more agents areadministered sequentially or simultaneously, they are considered to beadministered in combination for purposes of the present invention.

The IDO inhibitors of the present invention may be used in combinationwith at least one other (active) agent in any manner appropriate underthe circumstances. In one embodiment, treatment with the at least oneactive agent and at least one IDO inhibitor of the present invention ismaintained over a period of time. In another embodiment, treatment withthe at least one active agent is reduced or discontinued (e.g., when thesubject is stable), while treatment with an IDO inhibitor of the presentinvention is maintained at a constant dosing regimen. In a furtherembodiment, treatment with the at least one active agent is reduced ordiscontinued (e.g., when the subject is stable), while treatment with anIDO inhibitor of the present invention is reduced (e.g., lower dose,less frequent dosing or shorter treatment regimen). In yet anotherembodiment, treatment with the at least one active agent is reduced ordiscontinued (e.g., when the subject is stable), and treatment with theIDO inhibitor of the present invention is increased (e.g., higher dose,more frequent dosing or longer treatment regimen). In yet anotherembodiment, treatment with the at least one active agent is maintainedand treatment with the IDO inhibitor of the present invention is reducedor discontinued (e.g., lower dose, less frequent dosing or shortertreatment regimen). In yet another embodiment, treatment with the atleast one active agent and treatment with the IDO inhibitor of thepresent invention are reduced or discontinued (e.g., lower dose, lessfrequent dosing or shorter treatment regimen).

Oncology-Related Disorders.

The present invention provides methods for treating and/or preventing aproliferative condition, cancer, tumor, or precancerous disease,disorder or condition with an IDO inhibitor and at least one additionaltherapeutic agent, such as radiation, an immunomodulatory agent orchemotherapeutic agent, or diagnostic agent. Suitable immunomodulatoryagents that may be used in the present invention include CD40L, B7, andB7RP1; activating monoclonal antibodies (mAbs) to stimulatory receptors,such as, ant-CD40, anti-CD38, anti-ICOS, and 4-IBB ligand; dendriticcell antigen loading (in vitro or in vivo); anti-cancer vaccines such asdendritic cell cancer vaccines; cytokines/chemokines, such as, IL1, IL2,IL12, IL18, ELC/CCL19, SLC/CCL21, MCP-1, IL-4, IL-18, TNF, IL-15, MDC,IFNa/b, M-CSF, IL-3, GM-CSF, IL-13, and anti-IL-10; bacteriallipopolysaccharides (LPS); and immune-stimulatory oligonucleotides.

Examples of chemotherapeutic agents include, but are not limited to,alkylating agents such as thiotepa and cyclosphosphamide; alkylsulfonates such as busulfan, improsulfan and piposulfan; aziridines suchas benzodopa, carboquone, meturedopa, and uredopa; ethylenimines andmethylamelamines including altretamine, triethylenemelamine,trietylenephosphoramide, triethylenethiophosphaoramide andtrimethylolomelamime; nitrogen mustards such as chiorambucil,chlornaphazine, cholophosphamide, estramustine, ifosfamide,mechlorethamine, mechlorethamine oxide hydrochloride, melphalan,novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard;nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine,nimustine, ranimustine; antibiotics such as aclacinomysins, actinomycin,authramycin, azaserine, bleomycins, cactinomycin, calicheamicin,carabicin, caminomycin, carzinophilin, chromomycins, dactinomycin,daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin,epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins,mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin,puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin,tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such asmethotrexate and 5-fluorouracil (5-FU); folic acid analogs such asdenopterin, methotrexate, pteropterin, trimetrexate; purine analogs suchas fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine,5-FU; androgens such as calusterone, dromostanolone propionate,epitiostanol, mepitiostane, testolactone; anti-adrenals such asaminoglutethimide, mitotane, trilostane; folic acid replenisher such asfrolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinicacid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine;demecolcine; diaziquone; elformithine; elliptinium acetate; etoglucid;gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone;mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin;podophyllinic acid; 2-ethylhydrazide; procarbazine; razoxane; sizofiran;spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-trichlorotriethylamine; urethan; vindesine; dacarbazine;mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine;arabinoside (Ara-C); cyclophosphamide; thiotepa; taxoids, e.g.,paclitaxel and doxetaxel; chlorambucil; gemcitabine; 6-thioguanine;mercaptopurine; methotrexate; platinum and platinum coordinationcomplexes such as cisplatin and carboplatin; vinblastine; etoposide(VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine;vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin;xeloda; ibandronate; CPT11; topoisomerase inhibitors;difluoromethylornithine (DMFO); retinoic acid; esperamicins;capecitabine; and pharmaceutically acceptable salts, acids orderivatives of any of the above.

Chemotherapeutic agents also include anti-hormonal agents that act toregulate or inhibit hormonal action on tumors such as anti-estrogens,including for example tamoxifen, raloxifene, aromatase inhibiting4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, onapristone,and toremifene; and antiandrogens such as flutamide, nilutamide,bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptablesalts, acids or derivatives of any of the above. In certain embodiments,combination therapy comprises administration of a hormone or relatedhormonal agent.

Chemotherapeutic agents also include signal transduction inhibitors(STI). The term “signal transduction inhibitor” refers to an agent thatselectively inhibits one or more steps in a signaling pathway. Signaltransduction inhibitors (STIs) of the present invention include: (i)bcr/abl kinase inhibitors (e.g., GLEEVEC); (ii) epidermal growth factor(EGF) receptor inhibitors, including kinase inhibitors and antibodies;(iii) her-2/neu receptor inhibitors (e.g., HERCEPTIN); (iv) inhibitorsof Akt family kinases or the Akt pathway (e.g., rapamycin); (v) cellcycle kinase inhibitors (e.g., flavopiridol); and (vi) phosphatidylinositol kinase inhibitors.

Additional treatment modalities that may be used in combination with anIDO inhibitor include a cytokine or cytokine antagonist, such as IL-12,IFN, or anti-epidermal growth factor receptor, radiotherapy, amonoclonal antibody against another tumor antigen, a complex of amonoclonal antibody and toxin, a T-cell adjuvant, bone marrowtransplant, or antigen presenting cells (e.g., dendritic cell therapy).Vaccines (e.g., as a soluble protein or as a nucleic acid encoding theprotein) are also provided herein.

Cardiovascular Diseases.

The present invention provides methods for treating and/or preventingcertain cardiovascular- and/or metabolic-related diseases, disorders andconditions, as well as disorders associated therewith, with an IDOinhibitor and at least one additional therapeutic or diagnostic agent.

Examples of therapeutic agents useful in combination therapy for thetreatment of hypercholesterolemia (and atherosclerosis as well) includestatins (e.g., CRESTOR, LESCOL, LIPITOR, MEVACOR, PRAVACOL, and ZOCOR),which inhibit the enzymatic synthesis of cholesterol; bile acid resins(e.g., COLESTID, LO-CHOLEST, PREVALITE, QUESTRAN, and WELCHOL), whichsequester cholesterol and prevent its absorption; ezetimibe (ZETIA),which blocks cholesterol absorption; fibric acid (e.g., TRICOR), whichreduces triglycerides and may modestly increase HDL; niacin (e.g.,NIACOR), which modestly lowers LDL cholesterol and triglycerides; and/ora combination of the aforementioned (e.g., VYTORIN (ezetimibe withsimvastatin). Alternative cholesterol treatments that may be candidatesfor use in combination with the IDO inhibitors described herein includevarious supplements and herbs (e.g., garlic, policosanol, and guggul).The present invention encompasses pharmaceutically acceptable salts,acids or derivatives of any of the above.

Immune- and Inflammatory-Related Disorders.

The present invention provides methods for treating and/or preventingimmune- and/or inflammatory-related diseases, disorders and conditions,as well as disorders associated therewith, with an IDO inhibitor and atleast one additional therapeutic or diagnostic agent.

Examples of therapeutic agents useful in combination therapy include,but are not limited to, the following: non-steroidal anti-inflammatorydrug (NSAID) such as aspirin, ibuprofen, and other propionic acidderivatives (alminoprofen, benoxaprofen, bucloxic acid, carprofen,fenbufen, fenoprofen, fluprofen, flurbiprofen, indoprofen, ketoprofen,miroprofen, naproxen, oxaprozin, pirprofen, pranoprofen, suprofen,tiaprofenic acid, and tioxaprofen), acetic acid derivatives(indomethacin, acemetacin, alclofenac, clidanac, diclofenac,fenclofenac, fenclozic acid, fentiazac, fuirofenac, ibufenac, isoxepac,oxpinac, sulindac, tiopinac, tolmetin, zidometacin, and zomepirac),fenamic acid derivatives (flufenamic acid, meclofenamic acid, mefenamicacid, niflumic acid and tolfenamic acid), biphenylcarboxylic acidderivatives (diflunisal and flufenisal), oxicams (isoxicam, piroxicam,sudoxicam and tenoxican), salicylates (acetyl salicylic acid,sulfasalazine) and the pyrazolones (apazone, bezpiperylon, feprazone,mofebutazone, oxyphenbutazone, phenylbutazone). Other combinationsinclude cyclooxygenase-2 (COX-2) inhibitors.

Other active agents for combination include steroids such asprednisolone, prednisone, methylprednisolone, betamethasone,dexamethasone, or hydrocortisone. Such a combination may be especiallyadvantageous since one or more adverse effects of the steroid can bereduced or even eliminated by tapering the steroid dose required.

Additional examples of active agents that may be used in combinationsfor treating, for example, rheumatoid arthritis, include cytokinesuppressive anti-inflammatory drug(s) (CSAIDs); antibodies to, orantagonists of, other human cytokines or growth factors, for example,TNF, LT, IL-1β, IL-2, IL-6, IL-7, IL-8, IL-15, IL-16, IL-18, EMAP-II,GM-CSF, FGF, or PDGF.

Particular combinations of active agents may interfere at differentpoints in the autoimmune and subsequent inflammatory cascade, andinclude TNF antagonists such as chimeric, humanized or human TNFantibodies, REMICADE, anti-TNF antibody fragments (e.g., CDP870), andsoluble p55 or p75 TNF receptors, derivatives thereof, p75TNFRIgG(ENBREL.) or p55TNFR1gG (LENERCEPT), soluble IL-13 receptor (sIL-13),and also TNFα-converting enzyme (TACE) inhibitors; similarly, IL-1inhibitors (e.g., Interleukin-1-converting enzyme inhibitors) may beeffective. Other combinations include Interleukin 11, anti-P7s andp-selectin glycoprotein ligand (PSGL). Other examples of agents usefulin combination with the IDO inhibitors described herein includeinterferon-β1a (AVONEX); interferon-β1b (BETASERON); copaxone;hyperbaric oxygen; intravenous immunoglobulin; clabribine; andantibodies to, or antagonists of, other human cytokines or growthfactors (e.g., antibodies to CD40 ligand and CD80).

Immune Checkpoint Inhibitors.

The present invention contemplates the use of the inhibitors of IDOfunction described herein in combination with additional immunecheckpoint inhibitors.

The tremendous number of genetic and epigenetic alterations that arecharacteristic of all cancers provides a diverse set of antigens thatthe immune system can use to distinguish tumor cells from their normalcounterparts. In the case of T cells, the ultimate amplitude (e.g.,levels of cytokine production or proliferation) and quality (e.g., thetype of immune response generated, such as the pattern of cytokineproduction) of the response, which is initiated through antigenrecognition by the T-cell receptor (TCR), is regulated by a balancebetween co-stimulatory and inhibitory signals (immune checkpoints).Under normal physiological conditions, immune checkpoints are crucialfor the prevention of autoimmunity (i.e., the maintenance ofself-tolerance) and also for the protection of tissues from damage whenthe immune system is responding to pathogenic infection. The expressionof immune checkpoint proteins can be dysregulated by tumors as animportant immune resistance mechanism.

T cells have been the major focus of efforts to therapeuticallymanipulate endogenous antitumor immunity because of i) their capacityfor the selective recognition of peptides derived from proteins in allcellular compartments; ii) their capacity to directly recognize and killantigen-expressing cells (by CD8+ effector T cells; also known ascytotoxic T lymphocytes (CTLs)); and iii) their ability to orchestratediverse immune responses by CD4+ helper T cells, which integrateadaptive and innate effector mechanisms. In the clinical setting, theblockade of immune checkpoints—which results in the amplification ofantigen-specific T cell responses—has shown to be a promising approachin human cancer therapeutics.

T cell-mediated immunity includes multiple sequential steps, each ofwhich is regulated by counterbalancing stimulatory and inhibitorysignals in order to optimize the response. While nearly all inhibitorysignals in the immune response ultimately modulate intracellularsignaling pathways, many are initiated through membrane receptors, theligands of which are either membrane-bound or soluble (cytokines). Whileco-stimulatory and inhibitory receptors and ligands that regulate T-cellactivation are frequently not over-expressed in cancers relative tonormal tissues, inhibitory ligands and receptors that regulate T celleffector functions in tissues are commonly overexpressed on tumor cellsor on non-transformed cells associated with the tumor microenvironment.The functions of the soluble and membrane-bound receptor-ligand immunecheckpoints can be modulated using agonist antibodies (forco-stimulatory pathways) or antagonist antibodies (for inhibitorypathways). Thus, in contrast to most antibodies currently approved forcancer therapy, antibodies that block immune checkpoints do not targettumor cells directly, but rather target lymphocyte receptors or theirligands in order to enhance endogenous antitumor activity. [See Pardoll,(April 2012) Nature Rev. Cancer 12:252-64].

Examples of immune checkpoints (ligands and receptors), some of whichare selectively upregulated in various types of tumor cells, that arecandidates for blockade include PD1 (programmed cell death protein 1);PDL1 (PD1 ligand); BTLA (B and T lymphocyte attenuator); CTLA4(cytotoxic T-lymphocyte associated antigen 4); TIM3 (T-cell membraneprotein 3); LAG3 (lymphocyte activation gene 3); A2aR (adenosine A2areceptor A2aR); and Killer Inhibitory Receptors, which can be dividedinto two classes based on their structural features: i) killer cellimmunoglobulin-like receptors (KIRs), and ii) C-type lectin receptors(members of the type II transmembrane receptor family). Other lesswell-defined immune checkpoints have been described in the literature,including both receptors (e.g., the 2B4 (also known as CD244) receptor)and ligands (e.g., certain B7 family inhibitory ligands such B7-H3 (alsoknown as CD276) and B7-H4 (also known as B7-S1, B7x and VCTN1)). [SeePardoII, (April 2012) Nature Rev. Cancer 12:252-64].

The present invention contemplates the use of the inhibitors of IDOfunction described herein in combination with inhibitors of theaforementioned immune-checkpoint receptors and ligands, as well asyet-to-be-described immune-checkpoint receptors and ligands. Certainmodulators of immune checkpoints are currently available, whereas othersare in late-stage development. To illustrate, when it was approved forthe treatment of melanoma in 2011, the fully humanized CTLA4 monoclonalantibody ipilimumab (YERVOY; Bristol-Myers Squibb) became the firstimmune checkpoint inhibitor to receive regulatory approval in the US.Fusion proteins comprising CTLA4 and an antibody (CTLA4-Ig; abatcept(ORENCIA; Bristol-Myers Squibb)) have been used for the treatment ofrheumatoid arthritis, and other fusion proteins have been shown to beeffective in renal transplantation patients that are sensitized toEpstein Barr Virus. PD1 antibodies are also available for the treatmentof cancer, including for example nivolumab (Bristol-Myers Squibb) andpembroluzimab (Merck), and anti-PDL1 antibodies are also being evaluated(e.g., MPDL3280A (Roche)). Nivolumab (Opdivo®) has shown promise inpatients with melanoma, lung and kidney cancer, as well as multipleother malignancies.

In one aspect of the present invention, the claimed IDO inhibitors arecombined with an immuno-oncology agent that is (i) an agonist of astimulatory (including a co-stimulatory) receptor or (ii) an antagonistof an inhibitory (including a co-inhibitory) signal on T cells, both ofwhich result in amplifying antigen-specific T cell responses. Certain ofthe stimulatory and inhibitory molecules are members of theimmunoglobulin super family (IgSF). One important family ofmembrane-bound ligands that bind to co-stimulatory or co-inhibitoryreceptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1),B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6.Another family of membrane bound ligands that bind to co-stimulatory orco-inhibitory receptors is the TNF family of molecules that bind tocognate TNF receptor family members, which includes CD40 and CD40L,OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB),TRAIL/Apo2-L, TRAILR1/DR4, TRAILR2/DR5, TRAILR3, TRAILR4, OPG, RANK,RANKL, TWEAKR/Fn14, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LTβR,LIGHT, DcR3, HVEM, VEGI/TL1A, TRAMP/DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1,Lymphotoxin α/TNFβ, TNFR2, TNFα, LTβR, Lymphotoxin α 1β2, FAS, FASL,RELT, DR6, TROY, NGFR.

In another aspect, the immuno-oncology agent is a cytokine that inhibitsT cell activation (e.g., IL-6, IL-10, TGF-β, VEGF, and otherimmunosuppressive cytokines) or a cytokine that stimulates T cellactivation, for stimulating an immune response.

In one aspect, T cell responses can be stimulated by a combination ofthe claimed IDO inhibitors and one or more of (i) an antagonist of aprotein that inhibits T cell activation (e.g., immune checkpointinhibitors) such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, Galectin9, CEACAM-1, BTLA, CD69, Galectin-1, TIGIT, CD113, GPR56, VISTA, 2B4,CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4, and/or (ii) an agonist of aprotein that stimulates T cell activation such as B7-1, B7-2, CD28,4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70,CD27, CD40, DR3 and CD2. Other agents that can be combined with the IDOinhibitors of the present invention for the treatment of cancer includeantagonists of inhibitory receptors on NK cells or agonists ofactivating receptors on NK cells. For example, compounds herein can becombined with antagonists of KIR, such as lirilumab.

Yet other agents for combination therapies include agents that inhibitor deplete macrophages or monocytes, including but not limited to CSF-1Rantagonists such as CSF-1R antagonist antibodies including RG7155(WO11/70024, WO11/107553, WO11/131407, WO13/87699, WO13/119716,WO13/132044) or FPA-008 (WO11/140249; WO13169264; WO14/036357).

In another aspect, the claimed IDO inhibitors can be used with one ormore of agonistic agents that ligate positive costimulatory receptors,blocking agents that attenuate signaling through inhibitory receptors,antagonists, and one or more agents that increase systemically thefrequency of anti-tumor T cells, agents that overcome distinct immunesuppressive pathways within the tumor microenvironment (e.g., blockinhibitory receptor engagement (e.g., PD-L1/PD-1 interactions), depleteor inhibit Tregs (e.g., using an anti-CD25 monoclonal antibody (e.g.,daclizumab) or by ex vivo anti-CD25 bead depletion), or reverse/preventT cell anergy or exhaustion) and agents that trigger innate immuneactivation and/or inflammation at tumor sites.

In one aspect, the immuno-oncology agent is a CTLA-4 antagonist, such asan antagonistic CTLA-4 antibody. Suitable CTLA-4 antibodies include, forexample, YERVOY (ipilimumab) or tremelimumab.

In another aspect, the immuno-oncology agent is a PD-1 antagonist, suchas an antagonistic PD-1 antibody. Suitable PD-1 antibodies include, forexample, OPDIVO (nivolumab), KEYTRUDA (pembrolizumab/lambrolizumab), orMEDI-0680 (AMP-514; WO2012/145493). The immuno-oncology agent may alsoinclude pidilizumab (CT-011), though its specificity for PD-1 bindinghas been questioned. Another approach to target the PD-1 receptor is therecombinant protein composed of the extracellular domain of PD-L2(B7-DC) fused to the Fc portion of IgG1, called AMP-224

In another aspect, the immuno-oncology agent is a PD-L1 antagonist, suchas an antagonistic PD-L1 antibody. Suitable PD-L1 antibodies include,for example, MPDL3280A (RG7446; WO2010/077634), durvalumab (MEDI4736),BMS-936559 (WO2007/005874), and MSB0010718C (WO2013/79174).

In another aspect, the immuno-oncology agent is a LAG-3 antagonist, suchas an antagonistic LAG-3 antibody. Suitable LAG3 antibodies include, forexample, BMS-986016 (WO10/19570, WO14/08218), or IMP-731 or IMP-321(WO08/132601, WO09/44273).

In another aspect, the immuno-oncology agent is a CD137 (4-IBB) agonist,such as an agonistic CD137 antibody. Suitable CD137 antibodies include,for example, urelumab and PF-05082566 (WO12/32433).

In another aspect, the immuno-oncology agent is a GITR agonist, such asan agonistic GITR antibody. Suitable GITR antibodies include, forexample, BMS-986153, BMS-986156, TRX-518 (WO06/105021, WO09/009116) andMK-4166 (WO11/028683).

In another aspect, the immuno-oncology agent is an OX40 agonist, such asan agonistic OX40 antibody. Suitable OX40 antibodies include, forexample, MEDI-6383 or MEDI-6469.

In another aspect, the immuno-oncology agent is an OX40L antagonist,such as an antagonistic OX40 antibody. Suitable OX40L antagonistsinclude, for example, RG-7888 (WO06/029879).

In another aspect, the immuno-oncology agent is a CD40 agonist, such asan agonistic CD40 antibody. In yet another embodiment, theimmuno-oncology agent is a CD40 antagonist, such as an antagonistic CD40antibody. Suitable CD40 antibodies include, for example, lucatumumab ordacetuzumab.

In another aspect, the immuno-oncology agent is a CD27 agonist, such asan agonistic CD27 antibody. Suitable CD27 antibodies include, forexample, varlilumab.

In another aspect, the immuno-oncology agent is MGA271 (to B7H3)(WO11/109400).

The present invention encompasses pharmaceutically acceptable salts,acids or derivatives of any of the above.

Viral Diseases.

The present invention provides methods for treating and/or preventingviral diseases, disorders and conditions, as well as disordersassociated therewith, with an IDO inhibitor and at least one additionaltherapeutic or diagnostic agent (e.g., one or more other antiviralagents and/or one or more agents not associated with viral therapy).

Such combination therapy includes anti-viral agents targeting variousviral life-cycle stages and having different mechanisms of action,including, but not limiting to, the following: inhibitors of viraluncoating (e.g., amantadine and rimantidine); reverse transcriptaseinhibitors (e.g., acyclovir, zidovudine, and lamivudine); agents thattarget integrase; agents that block attachment of transcription factorsto viral DNA; agents (e.g., antisense molecules) that impact translation(e.g., fomivirsen); agents that modulate translation/ribozyme function;protease inhibitors; viral assembly modulators (e.g., rifampicin);antiretrovirals such as, for example, nucleoside analogue reversetranscriptase inhibitors (e.g., azidothymidine (AZT), ddl, ddC, 3TC,d4T); non-nucleoside reverse transcriptase inhibitors (e.g., efavirenz,nevirapine); nucleotide analogue reverse transcriptase inhibitors; andagents that prevent release of viral particles (e.g., zanamivir andoseltamivir). Treatment and/or prevention of certain viral infections(e.g., HIV) frequently entail a group (“cocktail”) of antiviral agents.

Other antiviral agents contemplated for use in combination with an IDOinhibitor include, but are not limited to, the following: abacavir,adefovir, amantadine, amprenavir, ampligen, arbidol, atazanavir,atripla, boceprevirertet, cidofovir, combivir, darunavir, delavirdine,didanosine, docosanol, edoxudine, emtricitabine, enfuvirtide, entecavir,famciclovir, fosamprenavir, foscarnet, fosfonet, ganciclovir,ibacitabine, imunovir, idoxuridine, imiquimod, indinavir, inosine,various interferons (e.g., peginterferon alfa-2a), lopinavir, loviride,maraviroc, moroxydine, methisazone, nelfinavir, nexavir, penciclovir,peramivir, pleconaril, podophyllotoxin, raltegravir, ribavirin,ritonavir, pyramidine, saquinavir, stavudine, telaprevir, tenofovir,tipranavir, trifluridine, trizivir, tromantadine, truvada, valaciclovir,valganciclovir, vicriviroc, vidarabine, viramidine, and zalcitabine.

The present invention encompasses pharmaceutically acceptable salts,acids or derivatives of any of the above.

Parasitic Disorders.

The present invention contemplates the use of the inhibitors of IDOfunction described herein in combination with antiparasitic agents. Suchagents include, but are not limited to, thiabendazole, pyrantel pamoate,mebendazole, praziquantel, niclosamide, bithionol, oxamniquine,metrifonate, ivermectin, albendazole, eflornithine, melarsoprol,pentamidine, benznidazole, nifurtimox, and nitroimidazole.

The skilled artisan is aware of other agents that may find utility forthe treatment of parasitic disorders.

The present invention encompasses pharmaceutically acceptable salts,acids or derivatives of any of the above.

Bacterial Infections.

Embodiments of the present invention contemplate the use of the IDOinhibitors described herein in combination with agents useful in thetreatment or prevention of bacterial disorders. Antibacterial agents canbe classified in various manners, including based on mechanism ofaction, based on chemical structure, and based on spectrum of activity.Examples of antibacterial agents include those that target the bacterialcell wall (e.g., cephalosporins and penicillins) or the cell membrane(e.g., polymyxins), or interfere with essential bacterial enzymes (e.g.,sulfonamides, rifamycins, and quinolines). Most antibacterial agentsthat target protein synthesis (e.g., tetracyclines and macrolides) arebacteriostatic, whereas agents such as the aminoglycoside arebactericidal. Another means of categorizing antibacterial agents isbased on their target specificity; “narrow-spectrum” agents targetspecific types of bacteria (e.g., Gram-positive bacteria such asStreptococcus), while “broad-spectrum” agents have activity against abroader range of bacteria. The skilled artisan is aware of types ofanti-bacterial agents that are appropriate for use in specific bacterialinfections.

The present invention encompasses pharmaceutically acceptable salts,acids or derivatives of the agents (and members of the classes ofagents) set forth above.

Dosing

The IDO inhibitors of the present invention may be administered to asubject in an amount that is dependent upon, for example, the goal ofadministration (e.g., the degree of resolution desired); the age,weight, sex, and health and physical condition of the subject to whichthe formulation is being administered; the route of administration; andthe nature of the disease, disorder, condition or symptom thereof. Thedosing regimen may also take into consideration the existence, nature,and extent of any adverse effects associated with the agent(s) beingadministered. Effective dosage amounts and dosage regimens can readilybe determined from, for example, safety and dose-escalation trials, invivo studies (e.g., animal models), and other methods known to theskilled artisan.

In general, dosing parameters dictate that the dosage amount be lessthan an amount that could be irreversibly toxic to the subject (themaximum tolerated dose (MTD)) and not less than an amount required toproduce a measurable effect on the subject. Such amounts are determinedby, for example, the pharmacokinetic and pharmacodynamic parametersassociated with ADME, taking into consideration the route ofadministration and other factors.

An effective dose (ED) is the dose or amount of an agent that produces atherapeutic response or desired effect in some fraction of the subjectstaking it. The “median effective dose” or ED50 of an agent is the doseor amount of an agent that produces a therapeutic response or desiredeffect in 50% of the population to which it is administered. Althoughthe ED50 is commonly used as a measure of reasonable expectance of anagent's effect, it is not necessarily the dose that a clinician mightdeem appropriate taking into consideration all relevant factors. Thus,in some situations the effective amount is more than the calculatedED50, in other situations the effective amount is less than thecalculated ED50, and in still other situations the effective amount isthe same as the calculated ED50.

In addition, an effective dose of the IDO inhibitors of the presentinvention may be an amount that, when administered in one or more dosesto a subject, produces a desired result relative to a healthy subject.For example, for a subject experiencing a particular disorder, aneffective dose may be one that improves a diagnostic parameter, measure,marker and the like of that disorder by at least about 5%, at leastabout 10%, at least about 20%, at least about 25%, at least about 30%,at least about 40%, at least about 50%, at least about 60%, at leastabout 70%, at least about 80%, at least about 90%, or more than 90%,where 100% is defined as the diagnostic parameter, measure, marker andthe like exhibited by a normal subject.

For administration of an oral agent, the compositions can be provided inthe form of tablets, capsules and the like containing from 1.0 to 1000milligrams of the active ingredient, particularly 1.0, 3.0, 5.0, 10.0,15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0,500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of the activeingredient.

In certain embodiments, the dosage of the desired IDO inhibitor iscontained in a “unit dosage form”. The phrase “unit dosage form” refersto physically discrete units, each unit containing a predeterminedamount of the IDO inhibitor, either alone or in combination with one ormore additional agents, sufficient to produce the desired effect. Itwill be appreciated that the parameters of a unit dosage form willdepend on the particular agent and the effect to be achieved.

Kits

The present invention also contemplates kits comprising an IDOinhibitor, and pharmaceutical compositions thereof. The kits aregenerally in the form of a physical structure housing variouscomponents, as described below, and may be utilized, for example, inpracticing the methods described above.

A kit can include one or more of the IDO inhibitors disclosed herein(provided in, e.g., a sterile container), which may be in the form of apharmaceutical composition suitable for administration to a subject. TheIDO inhibitors can be provided in a form that is ready for use (e.g., atablet or capsule) or in a form requiring, for example, reconstitutionor dilution (e.g., a powder) prior to administration. When the IDOinhibitors are in a form that needs to be reconstituted or diluted by auser, the kit may also include diluents (e.g., sterile water), buffers,pharmaceutically acceptable excipients, and the like, packaged with orseparately from the IDO inhibitors. When combination therapy iscontemplated, the kit may contain the several agents separately or theymay already be combined in the kit. Each component of the kit may beenclosed within an individual container, and all of the variouscontainers may be within a single package. A kit of the presentinvention may be designed for conditions necessary to properly maintainthe components housed therein (e.g., refrigeration or freezing).

A kit may contain a label or packaging insert including identifyinginformation for the components therein and instructions for their use(e.g., dosing parameters, clinical pharmacology of the activeingredient(s), including mechanism of action, pharmacokinetics andpharmacodynamics, adverse effects, contraindications, etc.). Labels orinserts can include manufacturer information such as lot numbers andexpiration dates. The label or packaging insert may be, e.g., integratedinto the physical structure housing the components, contained separatelywithin the physical structure, or affixed to a component of the kit(e.g., an ampule, tube or vial).

Labels or inserts can additionally include, or be incorporated into, acomputer readable medium, such as a disk (e.g., hard disk, card, memorydisk), optical disk such as CD- or DVD-ROM/RAM, DVD, MP3, magnetic tape,or an electrical storage media such as RAM and ROM or hybrids of thesesuch as magnetic/optical storage media, FLASH media or memory-typecards. In some embodiments, the actual instructions are not present inthe kit, but means for obtaining the instructions from a remote source,e.g., via the internet, are provided.

Experimental

The following Examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the present invention, and are not intended to limit thescope of what the inventors regard as their invention, nor are theyintended to represent that the experiments below were performed or thatthey are all of the experiments that may be performed. It is to beunderstood that exemplary descriptions written in the present tense werenot necessarily performed, but rather that the descriptions can beperformed to generate data and the like of a nature described therein.Efforts have been made to ensure accuracy with respect to numbers used(e.g., amounts, temperature, etc.), but some experimental errors anddeviations should be accounted for.

Unless indicated otherwise, parts are parts by weight, molecular weightis weight average molecular weight, temperature is in degrees Celsius (°C.), and pressure is at or near atmospheric. Standard abbreviations areused, including the following: wt=wildtype; bp=base pair(s);kb=kilobase(s); nt=nucleotides(s); aa=amino acid(s); s or sec=second(s);min=minute(s); h or hr=hour(s); ng=nanogram; μg=microgram; mg=milligram;g=gram; kg=kilogram; dl or dL=deciliter; μl or μL=microliter; ml ormL=milliliter; l or L=liter; μM=micromolar; mM=millimolar; M=molar;kDa=kilodalton; i.m.=intramuscular(ly); i.p.=intraperitoneal(ly); SC orSQ=subcutaneous(ly); QD=daily; BID=twice daily; QW=weekly; QM=monthly;HPLC=high performance liquid chromatography; BW=body weight; U=unit;ns=not statistically significant; PBS=phosphate-buffered saline;IHC=immunohistochemistry; DMEM=Dulbecco's Modification of Eagle'sMedium; EDTA=ethylenediaminetetraacetic acid.

Materials and Methods

The following general materials and methods were used, where indicated,or may be used in the Examples below:

Standard methods in molecular biology are described in the scientificliterature (see, e.g., Sambrook et al., Molecular Cloning, ThirdEdition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.(2001); and Ausubel et al., Current Protocols in Molecular Biology,Vols. 1-4, John Wiley and Sons, Inc. New York, N.Y. (2001), whichdescribes cloning in bacterial cells and DNA mutagenesis (Vol. 1),cloning in mammalian cells and yeast (Vol. 2), glycoconjugates andprotein expression (Vol. 3), and bioinformatics (Vol. 4)).

The scientific literature describes methods for protein purification,including immunoprecipitation, chromatography, electrophoresis,centrifugation, and crystallization, as well as chemical analysis,chemical modification, post-translational modification, production offusion proteins, and glycosylation of proteins (see, e.g., Coligan etal., Current Protocols in Protein Science, Vols. 1-2, John Wiley andSons, Inc., NY (2000)).

Software packages and databases for determining, e.g., antigenicfragments, leader sequences, protein folding, functional domains,glycosylation sites, and sequence alignments, are available (see, e.g.,GCG® Wisconsin Package (Accelrys, Inc., San Diego, Calif.); andDECYPHER® (TimeLogic Corp., Crystal Bay, Nev.).

The literature is replete with assays and other experimental techniquesthat can serve as a basis for evaluation of the compounds describedherein.

An IDO enzyme assay and cellular production of kynurenine (KYN) isdescribed in Sarkar, S. A. et al., Diabetes, 56:72-79 (2007). Briefly,all chemicals can be purchased from Sigma-Aldrich (St. Louis, Mo.)unless specified otherwise. Groups of 1,000 human islets can be culturedfor 24 h in 1 mL medium with cytokines, recovered by centrifugation for5 min at 800×g and sonicated in 150 μL PBS containing a proteaseinhibitor cocktail (Set 2; Calbiochem, EMD Biosciences, San Diego,Calif.). The sonicate can be centrifuged for 10 min at 10,000×g, and thesupernatant can be assayed in triplicate by incubating a 40 μl samplewith an equal volume of 100 mmol/L potassium phosphate buffer, pH 6.5,containing 40 mmol/L ascorbic acid (neutralized to pH 7.0), 100 μmol/Lmethylene blue, 200 μg/mL catalase, and 400 μmol/l L-Trp for 30 min at37° C. The assay can be terminated by the addition of 16 μL 30% (w/v)trichloroacetic acid (TCA) and further incubated at 60° C. for 15 min tohydrolyze N-formylkynurenine to KYN. The mixture can then be centrifugedat 12,000 rpm for 15 min, and KYN can be quantified by mixing equalvolume of supernatant with 2% (w/v) Ehrlich's reagent in glacial aceticacid in 96-well microtiter plate and reading the absorbance at 480 nmusing L-KYN as standard. Protein in the islet samples can be quantifiedby Bio-Rad Protein assay at 595 nm. For the detection of L-KYN in theislet culture supernatants, proteins can be precipitated with 5% (w/v)TCA and centrifuged at 12,000 rpm for 15 min, and determination of KYNin the supernatant with Ehrlich's reagent can be determined as describedabove. IL-4 (10 μg/mL; 500-2,000 units/mL) and 1-α-methyl Trp (1-MT; 40μmol/L) can be added to the incubation media as indicated. This assaycan also form the basis of a cell-based assay, and may be quantified viaLCMS/MS as an alternative to UV/Vis detection.

Western Blot Analyses.

Groups of 1,000-1,200 islets incubated for 24 h in Miami medium in thepresence of cytokines can be harvested and sonicated in PBS as above,and 50 μg protein samples can be electrophoresed on 10% SDS-PAGE gels.COS7 cells (0.6×10⁶ cells/60 mm3 petri dish) transfected with human-IDOplasmid (3 μg) or empty vector cells can be used as positive andnegative controls, respectively. Proteins can be transferredelectrophoretically onto polyvinylidine fluoride membranes by semidrymethod and blocked for 1 h with 5% (w/v) nonfat dry milk inTris-buffered saline and 0.1% Tween and then incubated overnight withanti-human mouse IDO antibody (1:500; Chemicon, Temecula, Calif.),phospho-STAT_(1α) p91, and STAT_(1α) p91 (1:500; Zymed, San Francisco,Calif.). Immunoreactive proteins can be visualized with ECL PLUS®Western blotting detection reagent (Amersham BioSciences,Buckinghamshire, U.K.) after incubation for 1 h with anti-mousehorseradish peroxidase-conjugated secondary antibody (JacksonImmunolabs, West Grove, Pa.).

Immunohistochemical Detection of IDO.

Islets can be fixed in 4% paraformaldehyde in PBS (Invitrogen) for 1 h,immobilized in molten 10% porcine skin gelatin blocks (37° C.), andembedded in optimal cutting temperature compound. Immunofluorescentstaining on islet tissue can be performed on 7 μm sections that werestained with antibodies raised against pancreatic duodenal homeobox 1(PDX1) and IDO. Antigen retrieval can be performed in a water bath for30 min in a buffer containing 10 mmol/1 Tris and 1 mmol/1 EDTA (pH 9.0)at 97° C. The sections can be blocked for 1 h with 5% normal goat serumin PBS. The tissues can then be reacted with mouse monoclonal anti-humanIDO antibody (1:20; Chemicon) and goat polyclonal anti-human PDX1antibody (1:2,000; which may be requested from Dr. Chris Wright, Schoolof Medicine, Vanderbilt, Tenn.) overnight at room temperature in a humidchamber. Secondary antibodies anti-goat (labeled with Cy3) andanti-mouse (labeled with Cy2) can be purchased from Jackson Immunolabsand can be used at a concentration of 1:200. The nuclei can be stainedwith Hoechst 33258 (Molecular Probes, Eugene, Oreg.). Images can beacquired by Intelligent Imaging System software from an Olympus 1X81inverted motorized microscope equipped with Olympus DSU (spinning diskconfocal) and Hamamatsu ORCA IIER monochromatic CCD camera.

Alternative means for evaluating the IDO inhibitors of the presentinvention are described in WO 2010/0233166 and are summarized hereafter.

Biochemical Assay.

cDNA clones for both human and mouse IDO have been isolated and verifiedby sequencing and are commercially available. In order to prepare IDOfor biochemical studies, C-terminal His-tagged IDO protein can beproduced in E. coli using the IPTG-inducible pET5a vector system andisolated over a nickel column. The yield of the partially purifiedprotein can be verified by gel electrophoresis and the concentrationestimated by comparison to protein standards. To assay IDO enzymaticactivity, a 96-well plate spectrophotometric assay for kynurenineproduction can be run following published procedures (see, e.g.,Littlejohn, T. K. et al., Prot. Exp. Purif., 19:22-29 (2000)). To screenfor IDO inhibitory activity, compounds can be evaluated at a singleconcentration of, for example, 200 μM against 50 ng of IDO enzyme in 100μL reaction volumes with tryptophan added at increasing concentrationsat, for example, 0, 2, 20, and 200 μM. Kynurenine production can bemeasured at 1 hour.

Cell-Based Assay.

COS-1 cells can be transiently transfected with a CMV promoter-drivenplasmid expressing IDO cDNA using Lipofectamine 2000 (Invitrogen) asrecommended by the manufacturer. A companion set of cells can betransiently transfected with TDO-expressing plasmid. Forty-eight hourspost-transfection, the cells can be apportioned into a 96-well format at6×10⁴ cells per well. The following day, the wells can be washed and newmedia (phenol red free) containing 20 μg/mL tryptophan can be addedtogether with inhibitor. The reaction can be stopped at 5 hours and thesupernatant removed and spectrophotometrically-assayed for kynurenine aspreviously described for the enzyme assay. To obtain initialconfirmation of IDO activity, compounds can be evaluated at a singleconcentration of, for example, 100 μM. More extensive dose-escalationprofiles can be collected for select compounds.

Pharmacodynamic and Pharmacokinetic Evaluation.

A pharmacodynamic assay can be based on measuring serum levels of bothkynurenine and tryptophan, and calculating the kynurenine/tryptophanratio provides an estimate of IDO activity that is independent ofbaseline tryptophan levels. Serum tryptophan and kynurenine levels canbe determined by HPLC analysis, and serum compound levels can optionallyalso be determined in the same HPLC run.

Compounds can be initially evaluated by challenging mice with LPS andthen subsequently administering a bolus dose of compound at the timethat the serum kynurenine level plateaus. As the kynurenine pool israpidly turned over with a half-life in serum of less than 10 minutes,pre-existing kynurenine is not expected to unduly mask the impact thatan IDO inhibitor has on kynurenine production. Each experiment caninclude non-LPS-exposed mice (to determine baseline kynurenine levelsagainst which to compare the other mice) and a set of LPS-exposed micedosed with vehicle alone (to provide a positive control for IDOactivation). Each compound can initially be evaluated in mice at asingle high i.p. bolus dose in the range of at least 100 mg/kg. Bloodcan be collected at defined time intervals (for example, 50 μL sample at5, 15, 30 min., 1, 2, 4, 6, 8, and 24 hr. following compoundadministration) for HPLC analysis of kynurenine and tryptophan levels(pharmacodynamic analysis) as well as for the level of compound(pharmacokinetic analysis). From the pharmacokinetic data the peak serumconcentration of compound achieved can be determined as well as theestimated rate of clearance. By comparing the level of compound in serumrelative to the kynurenine/tryptophan ratio at various time points, theeffective IC₅₀ for IDO inhibition in vivo can be roughly estimated.Compounds exhibiting efficacy can be evaluated to determine a maximumdose that achieves 100% IDO inhibition at the peak concentration.

HPLC/MS and Preparatory/Analytical HPLC Methods Employed inCharacterization or Purification of Examples

Analytical HPLC/MS was performed using the following methods:

Method A: Waters Acquity SDS using the following method: Linear Gradientof 2% to 98% Solvent B over 1.7 min; UV visualization at 220 nm; Column:BEH C18 2.1 mm×50 mm; 1.7 μm particle (Heated to Temp. 50° C.); Flowrate: 0.8 ml/min; Mobile Phase A: 100% Water, 0.05% TFA; Mobile Phase B:100% Acetonitrile, 0.05% TFA.

Method B: Column: Waters Acquity UPLC BEH C18, 2.1×50 mm, 1.7-μmparticles; Mobile Phase A: 5:95 acetonitrile:water with 10 mM ammoniumacetate; Mobile Phase B: 95:5 acetonitrile:water with 10 mM ammoniumacetate; Temperature: 50° C.; Gradient: 0-100% B over 3 minutes, then a0.75-minute hold at 100% B; Flow: 1.00 mL/min; Detection: UV at 220 nm.

Method C: Waters SFC-100 MS, Column: Chiral OJ-H 25×3 cm ID, 5 μm Flowrate: 100.0 mL/min, Mobile Phase: 80/20 CO₂/MeOH, Detector Wavelength:220 nm.

Method D: Aurora analytical SFC, Column: Chiral OJ-H 250×4.6 mm ID, 5μm, Flow rate: 2.0 mL/min, Mobile Phase: 80/20 CO₂/MeOH.

Method E: Berger Prep SFC, Column: Chiral AS 25×3 cm ID, 5 μm Flow rate:85.0 mL/min, Mobile Phase: 82/18 CO₂/MeOH w/ 0.1% DEA, DetectorWavelength: 220 nm.

Method F: Aurora analytical SFC, Column: Chiral AS 250×4.6 mm ID, 5 μm,Flow rate: 2.0 mL/min, Mobile Phase: 80/20 CO₂/MeOH w/ 0.1% DEA.

Method G: Berger Prep SFC, Column: Chiral AS 25×3 cm ID, 5 μm Flow rate:85.0 mL/min, Mobile Phase: 86/14 CO₂/MeOH, Detector Wavelength: 220 nm.

Method H: Aurora analytical SFC, Column: Chiral AS 250×4.6 mm ID, 5 μm,Flow rate: 2.0 mL/min, Mobile Phase: 85/15 CO₂/MeOH.

Method I: Column: Waters Acquity UPLC BEH C18, 2.1×50 mm, 1.7-μmparticles; Mobile Phase A: 5:95 acetonitrile:water with 0.1%trifluoroacetic acid; Mobile Phase B: 95:5 acetonitrile:water with 0.1%trifluoroacetic acid; Temperature: 50° C.; Gradient: 0-100% B over 3minutes, then a 0.75-minute hold at 100% B; Flow: 1.0 mL/min; Detection:UV at 220 nm.

Method J: Preparative Chromatographic Conditions: Instrument: BergerPrep SFC MGII (LVL-L4021 Lab) Column: Chiral IC 25×3 cm ID, 5 μm; Flowrate: 85.0 mL/min; Mobile Phase: 74/26 CO₂/MeOH; Detector Wavelength:220 nm.

Method K: Preparative Chromatographic Conditions: Instrument: BergerPrep SFC MGII (LVL-L4021 Lab) Column: Chiral IC 25×3 cm ID, 5 μm; Flowrate: 85.0 mL/min; Mobile Phase: 75/25 CO₂/MeOH hold for 18 minutes,60/40 CO₂/MeOH hold for 11 minutes, 75/25 CO₂/MeOH hold for 3 minutes;Detector Wavelength: 220 nm.

Method L: Preparative Conditions: Berger SFC MGII; Stage-1: Column:Chiral OD-H 25×3 cm ID, 5-μm particles; Mobile Phase: 82/18 CO₂/MeOH;Detector Wavelength: 220 nm; Flow: 85 mL/min. Stage-2: Chiral IF 25×3 cmID, 5-μm particles; Mobile Phase: 80/20 CO₂/MeOH; Detector Wavelength:220 nm; Flow: 85 mL/min. Analytical Conditions: Aurora analytical SFC;Stage-1: Column: Chiral OD-H 250×4.6 mm ID, 5 μm; Mobile Phase: 80/20CO₂/MeOH; Flow: 2.0 mL/min; Stage-2: Column: Chiral IF 250×4.6 mm ID, 5μm; Mobile Phase: 80/20 CO₂/MeOH; Flow: 2.0 mL/min. Tr corresponds tothe analytical condition.

Method M: Preparative Conditions: Berger SFC MGII; Stage-1: Column:Chiral OD-H 25×3 cm ID, 5-μm particles; Mobile Phase: 80/20 CO₂/MeOH;Detector Wavelength: 220 nm; Flow: 85 mL/min. Stage-2: Chiral IF 25×3 cmID, 5-μm particles; Mobile Phase: 80/20 CO₂/MeOH; Detector Wavelength:220 nm; Flow: 85 mL/min. Analytical Conditions: Aurora analytical SFC;Stage-1: Column: Chiral OD-H 250×4.6 mm ID, 5 μm; Mobile Phase: 80/20CO₂/MeOH; Flow: 2.0 mL/min; Stage-2: Column: Chiral IF 250×4.6 mm ID, 5μm; Mobile Phase: 80/20 CO₂/MeOH; Flow: 2.0 mL/min. Tr corresponds tothe analytical condition.

Method N: Preparative Conditions: Berger SFC MGII; Column: WHELK-O® 1KROMASIL® 25×3 cm ID, 5-μm particles; Mobile Phase: 80/20 CO₂/MeOH;Detector Wavelength: 220 nm; Flow: 85 mL/min. Analytical Conditions:Aurora analytical SFC; Column: WHELK-O® 1 KROMASIL® 250×4.6 mm ID, 5 μm;Mobile Phase: 80/20 CO₂/MeOH; Flow: 2.0 mL/min; Tr corresponds to theanalytical condition.

Method O: Preparative Conditions: Berger SFC MGII; Column: Chiral OJ25×3 cm ID, 5-μm; Mobile Phase: 90/10 CO₂/MeOH; Detector Wavelength: 220nm; Flow: 85 mL/min. Analytical Conditions: Aurora analytical SFC;Column: Chiral OJ 250×4.6 mm ID, 5 μm; Mobile Phase: 90/10 CO₂/MeOH;Flow: 2.0 mL/min; Tr corresponds to the analytical condition.

Method P: Preparative Conditions: Waters SFC-100 MS; Column: PHENOMENEX®LUX Cellulose-2 25×3 cm ID, 5 μm; Mobile Phase: 75/25 CO₂/MeOH; DetectorWavelength: 220 nm; Flow: 100 mL/min. Analytical Conditions: Auroraanalytical SFC; Column: PHENOMENEX® LUX Cellulose-2 250×4.6 mm ID, 5 μm;Mobile Phase: 75/25 CO₂/MeOH; Flow: 2.0 mL/min; Tr corresponds to theanalytical condition.

Method Q: Preparative Conditions: Berger SFC MGII; Column: Chiral AD25×3 cm ID, 5-μm; Mobile Phase: 80/20 CO₂/MeOH; Detector Wavelength: 220nm; Flow: 85 mL/min. Analytical Conditions: Aurora analytical SFC;Column: Chiral AD 250×4.6 mm ID, 5 μm; Mobile Phase: 80/20 CO₂/MeOH;Flow: 2.0 mL/min; Tr corresponds to the analytical condition.

Method R: Preparative Conditions: Berger SFC MGII; Column: Chiral AD25×3 cm ID, 5-μm; Mobile Phase: 87/13 CO₂/MeOH; Detector Wavelength: 220nm; Flow: 85 mL/min. Analytical Conditions: Aurora analytical SFC;Column: Chiral AD 250×4.6 mm ID, 5 μm; Mobile Phase: 85/15 CO₂/MeOH;Flow: 2.0 mL/min; Tr corresponds to the analytical condition.

Method S: Preparative Conditions: Berger SFC MGII; Column: Chiral IF25×3 cm ID, 5-μm; Mobile Phase: 75/25 CO₂/MeOH; Detector Wavelength: 220nm; Flow: 85 mL/min. Analytical Conditions: Aurora analytical SFC;Column: Chiral IF 250×4.6 mm ID, 5 μm; Mobile Phase: 70/30 CO₂/MeOH;Flow: 2.0 mL/min; Tr corresponds to the analytical condition.

Method T: Preparative Conditions: Waters SFC100-MS; Column: Chiral IC25×3 cm ID, 5-μm coupled to WHELK-O® R,R KROMASIL® 25×3 cm ID 5-μm;Mobile Phase: 70/30 CO₂/MeOH; Detector Wavelength: 220 nm; Flow: 100mL/min. Analytical Conditions: Aurora analytical SFC; Column: Chiral IC250×4.6 mm ID, 5 μm coupled to WHELK-O® R,R KROMASIL® 25×3 cm ID 5-μm;Mobile Phase: 70/30 CO₂/MeOH; Flow: 2.0 mL/min; Tr corresponds to theanalytical condition.

Method U: Preparative Conditions: Waters SFC100-MS; Column: Chiral OJ-H25×3 cm ID, 5-μm; Mobile Phase: 70/30 CO₂/MeOH; Detector Wavelength: 220nm; Flow: 100 mL/min. Analytical Conditions: Aurora analytical SFC;Column: Chiral OJ-H 250×4.6 mm ID, 5 μm; Mobile Phase: 70/30 CO₂/MeOH;Flow: 2.0 mL/min; Tr corresponds to the analytical condition.

Method V: Preparative Conditions: Berger SFC MGII; Column: ChiralWHELK-O® 25×3 cm ID, 5-μm; Mobile Phase: 80/20 CO₂/MeOH; DetectorWavelength: 220 nm; Flow: 85 mL/min. Analytical Conditions: Auroraanalytical SFC; Column: Chiral WHELK-O® 250×4.6 mm ID, 5 μm; MobilePhase: 80/20 CO₂/MeOH; Flow: 2.0 mL/min; Tr corresponds to theanalytical condition.

Method W: Preparative Conditions: Berger SFC MGH; Column: Chiral IC 25×3cm ID, 5-μm; Mobile Phase: 85/15 CO₂/MeOH; Detector Wavelength: 220 nm;Flow: 85 mL/min. Analytical Conditions: Aurora analytical SFC; Column:Chiral IC 250×4.6 mm ID, 5 μm; Mobile Phase: 85/15 CO₂/MeOH; Flow: 2.0mL/min; Tr corresponds to the analytical condition.

Method X: Preparative Conditions: Berger SFC MGII; Column: Chiral IC25×3 cm ID, 5-μm; Mobile Phase: 75/25 CO₂/MeOH w/0.1% diethylamine;Detector Wavelength: 220 nm; Flow: 85 mL/min. Analytical Conditions:Aurora analytical SFC; Column: Chiral IC 250×4.6 mm ID, 5 μm; MobilePhase: 75/25 CO₂/MeOH w/0.1% diethylamine; Flow: 2.0 mL/min; Trcorresponds to the analytical condition.

Method Y: Mobile Phase: 80/20 CO₂/MeOH/CAN 50/50; Flow: 2.0 mL/min; Trcorresponds: Preparative Conditions: Berger SFC MGII; Column: Chiral AD25×3 cm, 5-μm; Mobile Phase: 80/20 CO₂/MeOH/CAN 50/50; DetectorWavelength: 220 nm; Flow: 85 mL/min. Analytical Conditions: Auroraanalytical SFC; Column: Chiral AD 250×4.6 mm ID, 5 μm; to the analyticalcondition.

Method Z: Preparative Conditions: Berger SFC MGII; Column: Chiral IC25×3 cm, 5-μm; Mobile Phase: 83/17 CO₂/MeOH; Detector Wavelength: 220nm; Flow: 85 mL/min. Analytical Conditions: Aurora analytical SFC;Column: Chiral IC 250×4.6 mm ID, 5 μm; Mobile Phase: 80/20 CO₂/MeOH;Flow: 2.0 mL/min; Tr corresponds to the analytical condition.

Method AA: Preparative Conditions: Waters SFC100-MS; Column: Chiral AS-Hcoupled Chiral OJ-H 25×3 cm, 5-μm; Mobile Phase: 70/30 CO₂/MeOH;Detector Wavelength: 220 nm; Flow: 100 mL/min. Analytical Conditions:AGILENT® analytical SFC; Column: Chiral AS-H coupled to Chiral OJ-H250×4.6 mm ID, 5 μm; Mobile Phase: 70/30 CO₂/MeOH; Flow: 2.0 mL/min; Trcorresponds to the analytical condition.

Method AB: Waters Acquity SDS using the following method: LinearGradient of 2% to 98% Solvent B over 1.6 min; UV visualization at 220nm; Column: BEH C18 2.1 mm×50 mm; 1.7 μm particle (Heated to Temp. 50°C.); Flow rate: 1 ml/min; Mobile Phase A: 100% Water, 0.05% TFA; MobilePhase B: 100% Acetonitrile, 0.05% TFA.

Method AC: Preparative Conditions: Berger SFC MGII; Column: Chiral AD25×3 cm ID, 5-μm; Mobile Phase: 90/10 CO₂/MeOH; Detector Wavelength: 220nm; Flow: 85 mL/min. Analytical Conditions: Aurora analytical SFC;Column: Chiral AD 250×4.6 mm ID, 5 μm; Mobile Phase: 90/20 CO₂/MeOH;Flow: 2.0 mL/min; Tr corresponds to the analytical condition.

Method AD: Preparative Conditions: Berger SFC MGII; Column: Whelk-O1Kromasil 25×3 cm ID, 5-μm particles; Mobile Phase: 85/15 CO₂/MeOH;Detector Wavelength: 220 nm; Flow: 85 mL/min. Analytical Conditions:Aurora analytical SFC; Column: Whelk-O1 Kromasil 250×4.6 mm ID, 5 μm;Mobile Phase: 85/15 CO₂/MeOH; Flow: 2.0 mL/min; Tr corresponds to theanalytical condition.

Method AE: Preparative Conditions: Berger SFC MGII; Column: Whelk-O1Kromasil 25×3 cm ID, 5-μm particles; Mobile Phase: 75/25 CO₂/MeOH;Detector Wavelength: 220 nm; Flow: 85 mL/min. Analytical Conditions:Aurora analytical SFC; Column: Whelk-O1 Kromasil 250×4.6 mm ID, 5 μm;Mobile Phase: 75/25 CO₂/MeOH; Flow: 2.0 mL/min; Tr corresponds to theanalytical condition.

NMR Employed in Characterization of Examples

¹H NMR spectra (unless otherwise noted) were obtained with JEOL® orBruker FOURIER® transform spectrometers operating at 400 MHz or 500 MHz.

Spectral data are reported as chemical shift (multiplicity, number ofhydrogens, coupling constants in Hz) and are reported in ppm (δ units)relative to either an internal standard (tetramethyl silane=0 ppm) for¹H NMR spectra, or are referenced to the residual solvent peak (2.49 ppmfor CD₃SOCD₂H, 3.30 ppm for CD₂HOD, 1.94 for CHD₂CN, 7.26 ppm for CHCl₃,5.32 ppm for CDHCl₂). Abbreviations used in the description of NMRpeaks: “a”=apparent, “br. s.”=broad singlet.

EXAMPLES General Procedures

General Procedure A. Amide Bond Formation from Acid.

To a stirred solution of carboxylic acid (4.4 mmol) in dimethylformamide(DMF, 15 mL) was added aniline (6.6 mmol), diisopropylethylamine (1.53mL, 8.8 mmol) and1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (HATU) (2.00 g, 5.28 mmol). The resultingreaction mixture was stirred at rt for 3 h, at which point 3 M HCl (30mL) and CH₂Cl₂ (30 mL) were added. The layers were separated, and theaqueous layer was extracted with CH₂Cl₂ (2×30 mL). The combined organicextracts were dried over anhydrous sodium sulfate and concentrated underreduced pressure. The resulting crude residue was purified by silica gelchromatography to afford the desired product(s).

General Procedure B. Reaction Between Amines and Acyl Chlorides.

To a solution of the amine (1.1 equiv) and NEt₃ (5.0 equiv) in CH₂Cl₂(0.1 M) was added the acyl chloride (1.0 equiv). The resulting reactionmixture was stirred at rt for 15 min and then concentrated under reducedpressure. The crude reaction mixture was purified using silica gelchromatography (0% to 100% EtOAc in hexanes) to afford the desiredproduct.

Cis-4-phenylcyclohexane-1-carbaldehyde: Prepared according theliterature procedure (Fox, B. M. et al., J. Med. Chem., 57:3464-3483(2014)). The crude mixture was purified using silica gel chromatography(0% to 10% EtOAc in hexane) to afford the desired product as the firsteluting isomer.

Cis-(4-phenylcyclohexyl)methanol: To a solution ofcis-4-phenylcyclohexane-1-carbaldehyde (825 mg, 4.4 mmol) in THF (25 mL)and MeOH (7 mL) at rt was added NaBH₄ portionwise over 5 min. Theresulting mixture was stirred at rt for 45 min. Then HCl (1 M) was addeddropwise. The mixture was extracted with EtOAc (3×). The combinedorganic layers were washed with brine, dried over Na₂SO₄ andconcentrated under reduced pressure. The resulting crude mixture waspurified employing silica gel chromatography (0% to 25% EtOAc inhexanes) to afford the desired product.

Cis-(4-(iodomethyl)cyclohexyl)benzene: To a solution ofcis-(4-phenylcyclohexyl)methanol (2 g, 10.5 mmol), triphenylphosphine(3.3 g, 12.6 mmol) and imidazole (1.1 g, 15.8 mmol) in CH₂Cl₂ (70 mL) at0° C. was added iodine (3.5 g, 13.7 mmol). The mixture was warmed to rtand stirred at rt for 2 h. The mixture was diluted with CH₂Cl₂ andwashed with sodium thiosulfate (2 M). The organic layer was dried overanhydrous MgSO₄, filtered, and concentrated under reduced pressure. Thecrude reaction mixture was purified employing silica gel chromatography(0% to 25% EtOAc in hexanes) to afford the desired product as an oil (3g, 95%).

Cis-(4-(azidomethyl)cyclohexyl)benzene: To a solution ofcis-(4-(iodomethyl)cyclohexyl)benzene (2.6 g, 8.8 mmol) in DMF (44 mL)was added sodium azide (2.8 g, 43.8 mmol). The mixture was stirred at rtfor 2 h. Then more sodium azide (1.14 g, 17.5 mmol) was added and themixture was stirred at rt for 18 h. The mixture was diluted with Et₂Oand washed with water, 1 M LiCl (2×) and brine. The organic layers weredried over Na₂SO₄ and concentrated under reduced pressure to give thedesired product (1.5 g, 80%).

Cis-(4-phenylcyclohexyl)methanamine: To a solution ofcis-(4-(azidomethyl)cyclohexyl)benzene (1.5 g, 7.0 mmol) in THF (35 mL)was added triphenylphosphine (2.56 g, 9.8 mmol). The mixture was stirredat rt for 30 min and then water (0.83 mL) was added. The mixture wasstirred at rt for 24 h. The mixture was preabsorbed onto silica gel andpurified employing silica gel chromatography [0% to 5% (2 M NH₃ in MeOH)in CH₂Cl₂] to afford the desired product as an oil (1.2 g, 94%).

Example 1 Cis-4-cyano-N-((4-phenylcyclohexyl)methyl)benzamide

Prepared with General Procedure B employingcis-(4-phenylcyclohexyl)methanamine (19 mg, 0.1 mmol), 4-cyanobenzoylchloride (17 mg, 0.1 mmol), and NEt₃ (51 mg, 0.5 mmol) in CH₂Cl₂ (1 mL).Purified using silica gel chromatography (10% to 30% EtOAc in hexanes)to afford the desired product as a white solid. ¹H NMR (400 MHz; CDCl₃):δ 7.89-7.86 (m, 2H), 7.74-7.71 (m, 2H), 7.32-7.17 (m, 5H), 6.32-6.30 (m,1H), 3.58 (dd, J=7.7, 6.0 Hz, 2H), 2.66-2.59 (m, 1H), 2.06-2.00 (m, 1H),1.80-1.69 (m, 8H). m/z 319.3 (M+H⁺).

Example 2 Cis-3-cyano-N-((4-phenylcyclohexyl)methyl)benzamide

Prepared with General Procedure B employingcis-(4-phenylcyclohexyl)methanamine (19 mg, 0.1 mmol), 3-cyanobenzoylchloride (17 mg, 0.1 mmol), and NEt₃ (51 mg, 0.5 mmol) in CH₂Cl₂ (1 mL).Purified using silica gel chromatography (10% to 30% EtOAc in hexanes)to afford the desired product as a white solid. ¹H NMR (400 MHz; CDCl₃):δ 8.09-8.08 (m, 1H), 8.04 (dt, J=7.9, 1.5 Hz, 1H), 7.76 (dt, J=7.7, 1.3Hz, 1H), 7.56 (t, J=7.8 Hz, 1H), 7.32-7.17 (m, 5H), 6.49-6.46 (m, 1H),3.58 (dd, J=7.7, 6.0 Hz, 2H), 2.66-2.59 (m, 1H), 2.07-2.01 (m, 1H),1.80-1.68 (m, 8H). m/z 319.2 (M+H⁺).

Example 3 Cis-4-chloro-N-((4-phenylcyclohexyl)methyl)benzamide

Prepared with General Procedure B employingcis-(4-phenylcyclohexyl)methanamine (19 mg, 0.1 mmol), 4-chlorobenzoylchloride (19 mg, 0.1 mmol), and NEt₃ (51 mg, 0.5 mmol) in CH₂Cl₂ (1 mL).Purified using silica gel chromatography (0% to 20% EtOAc in hexanes) toafford the desired product as a white solid. ¹H NMR (400 MHz; CDCl₃): δ7.73-7.69 (m, 2H), 7.41-7.38 (m, 2H), 7.31-7.23 (m, 4H), 7.20-7.16 (m,1H), 3.55 (dd, J=7.7, 5.9 Hz, 2H), 2.63-2.59 (m, 1H), 2.04-1.98 (m, 1H),1.79-1.67 (m, 8H). m/z 328.2 (M+H⁺).

Example 4 Cis-3-chloro-N-((4-phenylcyclohexyl)methyl)benzamide

Prepared with General Procedure B employingcis-(4-phenylcyclohexyl)methanamine (19 mg, 0.1 mmol), 3-chlorobenzoylchloride (19 mg, 0.1 mmol), and NEt₃ (51 mg, 0.5 mmol) in CH₂Cl₂ (1 mL).Purified using silica gel chromatography (0% to 20% EtOAc in hexanes) toafford the desired product as a white solid. ¹H NMR (400 MHz; CDCl₃): δ7.76 (t, J=1.8 Hz, 1H), 7.64 (dt, J=7.7, 1.4 Hz, 1H), 7.47-7.44 (m, 1H),7.38-7.34 (m, 1H), 7.31-7.23 (m, 4H), 7.20-7.16 (m, 1H), 6.30-6.27 (m,1H), 3.56 (dd, J=7.7, 6.0 Hz, 2H), 2.64-2.58 (m, 1H), 2.04-1.99 (m, 1H),1.79-1.66 (m, 8H). m/z 328.2 (M+H⁺).

Example 5 Cis-4-fluoro-N-((4-phenylcyclohexyl)methyl)benzamide

Prepared with General Procedure B employingcis-(4-phenylcyclohexyl)methanamine (16 mg, 0.1 mmol), 4-fluorobenzoylchloride (19 mg, 0.1 mmol), and NEt₃ (51 mg, 0.5 mmol) in CH₂Cl₂ (1 mL).Purified using silica gel chromatography (0% to 20% EtOAc in hexanes) toafford the desired product as a white solid.

Example 6 Cis-4-chloro-N-((4-phenylcyclohexyl)methyl)benzenesulfonamide

Prepared in the manner of General Procedure B employingcis-(4-phenylcyclohexyl)methanamine (38 mg, 0.2 mmol),4-chlorobenzenesulfonyl chloride (42 mg, 0.2 mmol), and NEt₃ (101 mg,0.5 mmol) in CH₂Cl₂ (1 mL). Purified using silica gel chromatography (0%to 25% EtOAc in hexanes) to afford the desired product as a white solid.¹H NMR (400 MHz; CDCl₃): δ 7.87-7.83 (m, 2H), 7.52-7.48 (m, 2H),7.31-7.27 (m, 2H), 7.20-7.16 (m, 3H), 5.10 (t, J=6.2 Hz, 1H), 3.01 (dd,J=7.7, 6.3 Hz, 2H), 2.56 (dt, J=9.8, 5.0 Hz, 1H), 1.87-1.81 (m, 1H),1.68-1.52 (m, 8H). m/z 364.1 (M+H⁺).

Example 7 (4-Benzylpiperidin-1-yl)(4-chlorophenyl)methanone

To 4-chlorophenyl isocyanate (154 mg, 1.0 mmol) in Et₂O (5 mL) was added4-benzyl piperidine (193 mg, 1.1 mmol) The homogenous reaction mixtureproduced a precipitate over 15 min. The reaction mixture was cooled to0° C. and the solids were collected by filtration washing withadditional Et₂O (25 mL) to provide the desired product as a white solid.¹H NMR (400 MHz; CDCl₃): δ 7.31-7.28 (m, 4H), 7.25-7.21 (m, 3H),7.16-7.14 (m, 2H), 6.32 (s, 1H), 4.05-4.01 (m, 2H), 2.83 (td, J=12.9,2.1 Hz, 2H), 2.57 (d, J=6.9 Hz, 2H), 1.77-1.70 (m, 3H), 1.31-1.20 (m,2H). m/z 329.2 (M+H⁺).

Example 8 (4-Benzylpiperidin-1-yl)(3-chlorophenyl)methanone

To 3-chlorophenyl isocyanate (154 mg, 1.0 mmol) in Et₂O (5 mL) was added4-benzyl piperidine (193 mg, 1.1 mmol) The homogenous reaction mixtureproduced a precipitate over 15 min. The reaction mixture was cooled to0° C. and the solids were collected by filtration washing withadditional Et₂O (25 mL) to provide the desired product as a white solid.¹H NMR (400 MHz; CDCl₃): δ 7.47 (t, J=1.2 Hz, 1H), 7.29 (q, J=6.4 Hz,2H), 7.23-7.14 (m, 5H), 7.01-6.96 (m, 1H), 6.33 (s, 1H), 4.05-4.01 (m,2H), 2.83 (td, J=12.9, 2.1 Hz, 2H), 2.59-2.52 (m, 2H), 1.78-1.71 (m,3H), 1.31-1.21 (m, 2H). m/z 329.2 (M+H⁺).

Example 9 Cis-N-4-chloro-(2-(4-(4-methoxyphenyl)cyclohexyl)ethyl)aniline

9A. Ethyl 2-(4-(4-hydroxyphenyl)cyclohexylidene)acetateTriethylphosphonoacetate

(46.9 mL, 236 mmol) in THF (250 mL) was added dropwise over 1 hour at 0°C. to a solution of NaH (60% dispersion in oil, 11.8 g, 295 mmol) in THF(120 mL). The mixture was warmed to rt and stirred at rt for 1 h. In aseparate flask, a solution of 4-(4-hydroxyphenyl)cyclohexanone (37.5 g,197 mmol) in THF (250 mL) was added carefully to a mixture of NaH (60%dispersion in oil, 8.67 g, 216 mmol) in THF (100 mL) at 0° C. Themixture was stirred at rt for 2 hours. The mixture of the cyclohexanonewas added to the phosphonate mixture at 0° C. via cannulation. Themixture was warmed to rt and stirred at rt for 2 h. The mixture wasquenched by careful addition of ice and water (1 L) and subsequentlyextracted with ethyl acetate (3×500 mL) and the combined organics werethen washed with brine (1 L), dried over sodium sulfate, filtered, andconcentrated to provide ethyl2-(4-(4-hydroxyphenyl)cyclohexylidene)acetate in 97% yield as a whitesolid.

9B. Ethyl 2-(4-(4-hydroxyphenyl)cyclohexyl)acetate

To a solution ethyl 2-(4-(4-hydroxyphenyl)cyclohexylidene)acetate (9.74g, 35.8 mmol) in ethyl acetate was added Pd/C (0.974 g, 10 wt. %). Thereaction solution was sparged with a balloon of H₂ gas and stirredovernight under an atmosphere of hydrogen for 2 days. The reactionmixture was filtered through CELITE®, washing generously with ethylacetate, and concentrated under reduced pressure to afford the desiredproduct as a white crystalline solid in quantitative yield as a mixtureof diastereomers.

9C. Ethyl 2-(4-(4-methoxyphenyl)cyclohexyl)acetate

A solution of the product of Example 9B (20.0 g, 76.2 mmol, 1.0 equiv.)was dissolved in 770 mL of DMF. To this solution was added 6 mL (95mmol, 1.25 equiv.) of iodomethane followed by cesium carbonate (43.3 g,133 mmol, 1.75 equiv.). This mixture was then stirred for 16 hours untilstarting material was consumed as monitored by LCMS. The reaction wasthen quenched by cooling to 0° C. and subsequent addition of 1.35 L ofwater. The mixture was then extracted with ethyl acetate (3×500 mL) andthe combined organics were washed with brine (1 L) and dried over sodiumsulfate before filtration and concentration. The crude residue waspurified via column chromatography (5% ethyl acetate in hexanes) toafford the final compound as a clear oil in 69% yield. (R_(f)=0.5 in 10%ethyl acetate in hexanes).

9D. 2-(4-(4-Methoxyphenyl)cyclohexyl)acetic acid

Lithium hydroxide (1.58 g, 66.2 mmol) was added to water (8 mL). Theslurry was allowed to stand at rt for 30 min before filtering. Thefiltrate was added to a solution of the product of Example 9C (2.95 g,10.67 mmol) in EtOH (9 mL). The slurry was stirred at rt for 2 d anddiluted with water. The mixture was filtered and the solid was dilutedwith EtOAc and 1 M HCl. The layers were separated and the organic layerwas dried over sodium sulfate and concentrated under reduced pressure toprovide 2-(4-(4-methoxyphenyl)cyclohexyl)acetic acid.

9E and 9F.cis-N-(4-Chlorophenyl)-2-(4-(4-methoxyphenyl)cyclohexyl)acetamide andtrans-N-(4-Chlorophenyl)-2-(4-(4-methoxyphenyl)cyclohexyl)acetamide

Prepared with General Procedure A employing2-(4-(4-methoxyphenyl)cyclohexyl)acetic acid (product of Example 9D, 124mg, 0.5 mmol), 4-chloroaniline (97 mg, 0.75 mmol), HATU (435 mg, 0.75mmol), and ^(i)Pr₂NEt (323 mg, 2.5 mmol) in DMF (1.0 mL). Purificationusing silica gel chromatography (0% to 25% EtOAc in hexanes) affordedcis-N-(4-chlorophenyl)-2-(4-(4-methoxyphenyl)cyclohexyl)acetamide(Example 9E), a white solid, as the first eluting isomer andtrans-N-(4-chlorophenyl)-2-(4-(4-methoxyphenyl)cyclohexyl)acetamide(Example 9F) as the second eluting isomer.

cis-N-(4-Chlorophenyl)-2-(4-(4-methoxyphenyl)cyclohexyl)acetamide

¹H NMR (400 MHz; CDCl₃): δ 7.49-7.45 (m, 2H), 7.29-7.26 (m, 2H),7.17-7.15 (m, 3H), 6.87-6.83 (m, 2H), 3.79 (s, 3H), 2.63-2.55 (m, 1H),2.45-2.37 (m, 3H), 1.77-1.64 (m, 8H). m/z 358.2 (M+H⁺).

Example 9 Cis-N-4-chloro-(2-(4-(4-methoxyphenyl)cyclohexyl)ethyl)aniline

To a solution ofcis-N-(4-chlorophenyl)-2-(4-(4-methoxyphenyl)cyclohexyl)acetamide (33mg, 0.092 mmol) in THF (0.5 mL) at rt was added Borane tetrahydrofurancomplex solution (0.5 mL, 0.5 mmol, 1 M in THF). The resulting mixturewas stirred for 2.5 h at rt at which point aqueous HCl (1 M) was addedand the mixture was stirred at rt for 30 min. Evolution of gas wasobserved. The mixture was basified with sat. Na₂CO₃ and extracted withCH₂Cl₂ (2×). The combined organic layers were dried over anhydrousNa₂SO₄, filtered, and concentrated under reduced pressure. The resultingcrude mixture was purified employing silica gel chromatography (0% to10% EtOAc in hexanes) to afford the desired product. ¹H NMR (400 MHz,CDCl₃) δ 7.19-7.09 (m, 4H), 6.90-6.81 (m, 2H), 6.56 (d, J=8.7 Hz, 2H),3.80 (s, 3H), 3.17-3.07 (m, 2H), 2.55 (s, 1H), 1.85 (s, 1H), 1.80-1.63(m, 10H). m/z 344.2 (M+H⁺).

Example 10Trans-N-4-chloro-(2-(4-(4-methoxyphenyl)cyclohexyl)ethyl)aniline

Prepared using the procedure from the previous example employing 73 mgof trans-N-(4-chlorophenyl)-2-(4-(4-methoxyphenyl)cyclohexyl)acetamide.Purified using silica gel chromatography (0% to 10% EtOAc in hexanes) toafford the desired product. ¹H NMR (400 MHz, CDCl₃) δ 7.17-7.09 (m, 4H),6.89-6.81 (m, 2H), 6.58-6.51 (m, 2H), 3.80 (s, 3H), 3.18-3.09 (m, 2H),2.45 (t, J=12.3 Hz, 1H), 1.90 (d, J=12.3 Hz, 4H), 1.67-1.47 (m, 4H),1.44 (dd, J=17.5, 7.8 Hz, 2H), 1.14 (dd, J=22.1, 12.0 Hz, 2H). m/z 344.2(M+H⁺).

Example 11 (+/−)-Cis-3-phenylcyclopentyl(4-chlorophenyl)carbamate

11A. (+/+Cis-3-phenylcyclopentan-1-ol

3-Phenylcyclopentan-1-one was prepared as previously described(Yamamoto, T. et al., J. Organomet. Chem., 694:1325-1332 (2009)). To asolution of 3-phenylcyclopentan-1-one (1.0 g, 6.2 mmol) in 30 mL ofmethanol cooled to 0° C. was added NaBH₄ (0.27 g, 7.2 mmol). The icebath was removed, and the reaction was allowed to warm to rt and stirredfor 3 h. The reaction was quenched with 1 M HCl and diluted with EtOAc(30 mL), and the layers were separated. The organic extracts were driedover anhydrous MgSO₄, filtered, and concentrated under reduced pressure.The resulting crude mixture of ˜1.6:1 cis: trans alcohols was purifiedusing silica gel chromatography (15% EtOAc in pentane) to afford thedesired cis-3-phenylcyclopentan-1-ol as a colorless oil (118 mg, 12%).¹H NMR (400 MHz; CDCl₃): δ 7.16-7.09 (m, 5H), 4.43-4.40 (m, 1H),3.06-3.00 (m, 1H), 2.70 (br s, 1H), 2.53-2.43 (m, 1H), 2.06-1.63 (m,5H).

Example 11 (+/−)-Cis-3-phenylcyclopentyl(4-chlorophenyl)carbamate

To a solution of cis-3-phenylcyclopentan-1-ol (150 mg, 0.95 mmol) inCH₂CH₂ was added 4-chlorophenyl isocyanate (150 mg, 0.95 mmol). After 5min, the reaction mixture was concentrated under reduced pressure andtriturated with diethyl ether (10 mL). The mixture was filtered, rinsingwith diethyl ether, to afford the desired product as a white solid. ¹HNMR (400 MHz; DMSO-d₆): δ 9.79 (br s, 1H), 7.49 (d, J=7.5 Hz, 2H),7.33-7.14 (m, 7H), 5.09-5.04 (m, 1H), 3.13-3.02 (m, 1H), 2.60-2.48 (m,1H), 2.08-1.61 (m, 6H).

Example 12 Cis-(4-phenylcyclohexyl)methyl(4-fluorophenyl)carbamate

To a solution of cis-(4-phenylcyclohexyl)methanol (200 mg, 1.05 mmol) indiethyl ether (5 mL) was added 4-fluorophenyl isocyanate (144 mg, 1.05mmol). Upon consumption of the starting materials, the resultingsolution was concentrated to provide a white solid, which was trituratedin diethyl ether (3 mL) and filtered to afford the desired product as awhite solid. ¹H NMR (400 MHz; DMSO-d₆): δ 9.65 (br s, 1H), 7.48-7.42 (m,2H), 7.18-7.06 (m, 7H), 4.20 (d, J=9.5 Hz, 2H), 2.61-2.51 (m, 1H),2.07-2.01 (m, 1H), 1.77-1.57 (m, 8H).

Example 13

Trans-1-(4-fluorophenyl)-3-(4-phenylcyclohexyl)urea

To a stirred solution of trans-4-phenylcyclohexylamine (Combi-Blocks,San Diego, Calif.) (50 mg, 0.3 mmol) in diethyl ether was added4-fluorophenyl isocyanate (0.032 mL, 0.3 mmol) at rt. After stirring for30 min the voluminous white precipitate was isolated by vacuumfiltration to yield the desired product. ¹H NMR (400 MHz, CDCl₃) δ7.35-7.14 (m, 7H), 7.07-6.97 (m, 2H), 6.00 (s, 1H), 4.41-4.30 (m, 1H),3.83-3.65 (m, 1H), 2.56-2.39 (m, 1H), 2.21-2.10 (m, 2H), 1.99-1.88 (m,2H), 1.72-1.46 (m, 2H), 1.37-1.07 (m, 2H).

Example 14 Trans-1-(4-chlorophenyl)-3-(4-phenylcyclohexyl)urea

To a stirred solution of trans-4-phenylcyclohexylamine (50 mg, 0.3 mmol)in diethyl ether was added 4-chlorophenyl isocyanate (44 mg, 0.3 mmol)at rt. After stirring for 30 min the voluminous white precipitate wasisolated by vacuum filtration to yield the desired product. ¹H NMR (400MHz, CDCl₃) δ 7.38-7.12 (m, 9H), 6.05 (s, 1H), 4.39 (d, J=7.4 Hz, 1H),3.88-3.61 (m, 1H), 2.56-2.40 (m, 1H), 2.28-2.12 (m, 2H), 2.00-1.89 (m,2H), 1.71-1.59 (m, 2H), 1.36-1.16 (m, 2H).

Example 15 Trans-1-(3-chlorophenyl)-3-(4-phenylcyclohexyl)urea

To a stirred solution of trans-4-phenylcyclohexylamine (50 mg, 0.3 mmol)in diethyl ether (1.4 mL) was added 3-chlorophenyl isocyanate (0.035 mL,0.3 mmol) at rt. After stirring for 30 min the voluminous whiteprecipitate was isolated by vacuum filtration and concentrated underreduced pressure to yield the desired product. ¹H NMR (400 MHz, CDCl₃) δ7.42 (t, J=2.0 Hz, 1H), 7.34-7.11 (m, 7H), 7.08-7.02 (m, 1H), 6.14 (s,1H), 4.47 (s, 1H), 3.85-3.62 (m, 1H), 2.61-2.39 (m, 1H), 2.27-2.12 (m,2H), 2.02-1.89 (m, 2H), 1.74-1.57 (m, 2H), 1.39-1.19 (m, 2H).

Example 16 Trans-2-(3-chlorophenyl)-N-(4-phenylcyclohexyl)acetamide

Prepared according to General Procedure A usingtrans-4-phenylcyclohexylamine and 3-chlorophenylacetic acid. Purified bysilica gel chromatography (0% to 50% ethyl acetate in hexanes) whichafforded the desired product as white solid. ¹H NMR (400 MHz, CDCl₃) δ7.36-7.12 (m, 9H), 5.21 (d, J=7.9 Hz, 1H), 3.84 (tdt, J=12.0, 8.1, 4.0Hz, 1H), 3.53 (s, 2H), 2.51-2.37 (m, 1H), 2.13-1.99 (m, 2H), 1.99-1.85(m, 2H), 1.67-1.49 (m, 2H), 1.29-1.09 (m, 2H). m/z 328.2 (M+H⁺).

Example 17 Cis-1-(4-chlorophenyl)-3-(4-phenylcyclohexyl)urea

To a stirred solution of cis-4-phenylcyclohexylamine (Li, G. et al.,Bioorg. Med. Chem. Lett., 18:1146-1150 (2008)) (60 mg, 0.34 mmol) indiethyl ether (1.4 mL) was added 4-chlorophenyl isocyanate (53 mg, 0.34mmol) at rt. After stirring for 30 min the voluminous white precipitatewas isolated by vacuum filtration and concentrated under reducedpressure to yield the desired product. ¹H NMR (400 MHz, CDCl₃) δ7.37-7.09 (m, 9H), 6.28 (s, 1H), 4.88 (d, J=7.2 Hz, 1H), 4.17-4.03 (m,1H), 2.68-2.49 (m, 1H), 1.98-1.87 (m, 2H), 1.87-1.69 (m, 4H), 1.65-1.50(m, 2H).

Example 18 Cis-1-(3-chlorophenyl)-3-(4-phenylcyclohexyl)urea

To a stirred solution of cis-4-phenylcyclohexylamine (60 mg, 0.36 mmol)in diethyl ether (1.4 mL) was added 3-chlorophenyl isocyanate (0.042 mL,0.34 mmol) at rt. After stirring for 30 min the voluminous whiteprecipitate was isolated by vacuum filtration and concentrated underreduced pressure to yield the desired product. ¹H NMR (400 MHz, CDCl₃) δ7.47-7.42 (m, 1H), 7.35-7.14 (m, 7H), 7.04 (dt, J=7.5, 1.7 Hz, 1H), 6.37(s, 1H), 4.98 (d, J=6.6 Hz, 1H), 4.21-4.02 (m, 1H), 2.68-2.50 (m, 1H),2.00-1.87 (m, 2H), 1.88-1.67 (m, 4H), 1.67-1.49 (m, 2H).

Example 19 Cis-2-(4-chlorophenyl)-N-(4-phenylcyclohexyl)acetamide

Prepared according to General Procedure A usingcis-4-phenylcyclohexylamine and 4-chlorophenylacetic acid (Li, G. etal., Bioorg. Med. Chem. Lett., 18:1146-1150 (2008)). Purified by silicagel chromatography (0% to 50% ethyl acetate in hexanes) which affordedthe desired product as white solid. ¹H NMR (400 MHz, CDCl₃) δ 7.43-7.17(m, 7H), 7.07 (dd, J=7.5, 0.8 Hz, 2H), 5.86 (s, 1H), 4.25-4.05 (m, 1H),3.60 (s, 2H), 2.60-2.46 (m, 1H), 1.89-1.55 (m, 6H), 1.42-1.22 (m, 2H).m/z 328.2 (M+H⁺).

General Procedure C: Reaction Between Esters and Anilines.

To a solution of the aniline (2.0 equiv) in THF (0.25 M) at 0° C. wasadded a solution of ^(i)PrMgCl (2.0 equiv, 2 M in THF). The resultingsolution was warmed to rt and stirred for 5 min at which point the ester(1.0 equiv) was added dropwise. The resulting reaction mixture wasstirred at rt for 8 h and was poured into water. EtOAc was added, andthe layers were separated. The aqueous layer was extracted with EtOAc(3×). The combined organic extracts were dried over anhydrous MgSO₄,filtered, and concentrated under reduced pressure. The crude reactionmixture was purified using silica gel chromatography (0% to 100% EtOAcin hexanes) to afford the desired product(s).

Example 20 Cis-4-benzyl-N-(4-chlorophenyl)cyclohexane-1-carboxamide

Prepared using General Procedure C employing ethyl4-benzylcyclohexane-1-carboxylate (250 mg, 1.0 mmol), which can beprepared by methods shown in WO2005080317A2, and 4-chloroaniline (191mg, 1.5 mmol). Purification using silica gel chromatography (0% to 50%EtOAc in hexanes) afforded the desired product. ¹H NMR (400 MHz; CDCl₃):δ 7.49 (d, J=8.8 Hz, 2H), 7.28 (d, J=8.7 Hz, 2H), 7.23-7.10 (m, 5H),2.62 (d, J=7.7 Hz, 2H), 2.48-2.37 (m, 1H), 2.04-1.93 (m, 2H), 1.88-1.82(m, 2H), 1.74-1.43 (m, 4H), 1.10-0.93 (m, 1H); m/z 328.1 (M+H⁺).

Example 21 Trans-4-benzyl-N-(4-chlorophenyl)cyclohexane-1-carboxamide

Further elution from the column in the previous example afforded thedesired product as the second eluting isomer. ¹H NMR (400 MHz; CDCl₃): δ7.46 (d, J=8.8 Hz, 2H), 7.31-7.26 (m, 3H), 7.26-7.24 (m, 1H), 7.22-7.11(m, 4H), 2.52 (d, J=7.1 Hz, 2H), 2.20-2.10 (m, 1H), 1.97 (d, J=11.4 Hz,2H), 1.86-1.73 (m, 2H), 1.57-1.45 (m, 3H), 1.38-1.24 (m, 2H); m/z 328.1(M+H⁺).

Example 22 Cis-4-benzyl-N-(4-cyanophenyl)cyclohexane-1-carboxamide

Prepared using General Procedure C employing ethyl4-benzylcyclohexane-1-carboxylate (250 mg, 1.0 mmol) and4-aminobenzonitrile (177 mg, 1.5 mmol). Purification using silica gelchromatography (0% to 50% EtOAc in hexanes) afforded the desiredproduct. ¹H NMR (400 MHz; CDCl₃): δ 7.73-7.63 (m, 2H), 7.63-7.55 (m,2H), 7.51 (s, 1H), 7.32-7.24 (m, 2H), 7.23-7.10 (m, 3H), 2.61 (d, J=7.6Hz, 2H), 2.49-2.45 (m, 1H), 2.04-1.91 (m, 1H), 1.89-1.77 (m, 1H),1.74-1.46 (m, 7H); m/z 319.2 (M+H⁺).

Example 23 Trans-4-benzyl-N-(4-cyanophenyl)cyclohexane-1-carboxamide

Further elution from the column in the previous example afforded thedesired product as the second eluting isomer. ¹H NMR (400 MHz; CDCl₃): δ7.73-7.63 (m, 2H), 7.63-7.55 (m, 2H)), 7.46 (s, 1H), 7.32-7.24 (m, 2H),7.23-7.10 (m, 3H), 2.52 (d, J=7.1 Hz, 2H), 2.21 (tt, J=12.1, 3.4 Hz,1H), 2.04-1.91 (m, 2H), 1.89-1.77 (m, 2H), 1.74-1.46 (m, 3H), 1.03 (qd,J=13.2, 3.5 Hz, 2H).; m/z 319.2 (M+H⁺).

Example 24 Cis-4-benzyl-N-(4-fluorophenyl)cyclohexane-1-carboxamide

Prepared using General Procedure C employing ethyl4-benzylcyclohexane-1-carboxylate (250 mg, 1.0 mmol) and 4-fluoroaniline(0.15 mL, 1.5 mmol). Purification using silica gel chromatography (0% to50% EtOAc in hexanes) afforded the desired product. ¹H NMR (400 MHz;CDCl₃): δ 7.48 (ddd, J=10.5, 6.9, 4.8 Hz, 2H), 7.34-7.11 (m, 5H),7.09-6.94 (m, 3H), 2.62 (d, J=7.6 Hz, 2H), 2.48-2.36 (m, 1H), 2.04-1.92(m, 2H), 1.88-1.76 (m, 1H), 1.74-1.40 (m, 5H), 1.12-0.94 (m, 1H); m/z312.2 (M+H⁺).

Example 25 Trans-4-benzyl-N-(4-fluorophenyl)cyclohexane-1-carboxamide

Further elution from the column in the previous example afforded thedesired product as the second eluting isomer. ¹H NMR (400 MHz; CDCl₃): δ7.46 (dd, J=9.0, 4.8 Hz, 2H), 7.33-7.11 (m, 5H), 7.10-6.95 (m, 3H), 2.52(d, J=7.0 Hz, 2H), 2.20-2.11 (m, 1H), 1.97 (d, J=10.4 Hz, 2H), 1.84 (d,J=13.0 Hz, 2H), 1.61-1.44 (m, 3H), 1.00 (dd, J=29.4, 10.9 Hz, 2H); m/z312.2 (M+H⁺).

Example 26 Cis-4-benzyl-N-(4-methoxyphenyl)cyclohexane-1-carboxamide

Prepared using General Procedure C employing ethyl4-benzylcyclohexane-1-carboxylate (250 mg, 1.0 mmol) and4-methoxyaniline (185 mg, 1.5 mmol). Purification using silica gelchromatography (0% to 50% EtOAc in hexanes) afforded the desiredproduct. ¹H NMR (400 MHz; CDCl₃): δ 7.46-7.38 (m, 2H), 7.33-7.23 (m,2H), 7.22-7.09 (m, 4H), 6.92-6.80 (m, 2H), 3.79 (s, 3H), 2.62 (d, J=7.6Hz, 2H), 2.48-2.35 (m, 1H), 2.05-1.95 (m, 2H), 1.86-1.75 (m, 1H),1.70-1.63 (m, 2H), 1.65-1.48 (m, 4H); m/z 324.2 (M+H⁺).

Example 27 Trans-4-benzyl-N-(4-methoxyphenyl)cyclohexane-1-carboxamide

Further elution from the column in the previous example afforded thedesired product as the second eluting isomer. ¹H NMR (400 MHz; CDCl₃): δ7.40 (d, J=9.0 Hz, 2H), 7.29 (d, J=7.0 Hz, 2H), 7.23-7.09 (m, 3H), 7.01(s, 1H), 6.84 (d, J=8.9 Hz, 2H), 3.78 (s, 3H), 2.52 (d, J=6.9 Hz, 2H),2.15 (tt, J=12.2, 3.5 Hz, 1H), 1.98 (d, J=11.2 Hz, 2H), 1.83 (d, J=13.6Hz, 2H), 1.55-1.49 (m, 3H), 1.04 (qd, J=13.3, 3.3 Hz, 2H); m/z 324.2(M+H⁺).

Example 29 N-Benzyl-2-(4-(4-methoxyphenyl)cyclohexyl)acetamide

To solution of 2-(4-(4-methoxyphenyl)cyclohexyl)acetic acid (product of9D, 152 mg, 0.61 mmol) in CH₂Cl₂ (1.2 mL) at rt was added oxalylchloride (63 μL, 0.73 mmol) and one drop of DMF. Evolution of gas wasobserved and the mixture turned yellow in color. The mixture was stirredat rt for 1 h and then concentrated under reduced pressure. The residuewas dissolved in CH₂Cl₂ (1.2 mL) and benzyl amine (67 μL, 0.61 mmol) andtriethylamine (85 μL, 0.61 mmol) were added at rt. A white precipitateformed and more triethylamine (170 μL, 1.22 mmol) and CH₂Cl₂ (1.2 mL)were added. The homogenous mixture was stirred at rt for 3 h. Themixture was concentrated under reduced pressure. The residue wasdissolved in EtOAc and washed with sat. NaHCO₃ and brine. The organiclayer was dried over sodium sulfate and concentrated under reducedpressure. The residue was purified using silica gel chromatography (35%EtOAc in hexanes) to afford a mixture of isomers. The residue wasrecrystallized from heptane/IPA to afford the desired product as whitesolid as a 2:1 mixture of trans:cis isomers. m/z 338.3 (M+H⁺).

Example 30 Cis-N-benzyl-2-(4-(4-methoxyphenyl)cyclohexyl)acetamide

The mother liquors from the previous example were concentrated underreduced pressure to give the desired product. ¹H NMR (400 MHz, CDCl₃) δ7.45-7.23 (m, 5H), 7.20-7.07 (m, 2H), 6.92-6.71 (m, 2H), 5.80 (s, 1H),4.46 (d, J=5.7 Hz, 2H), 3.79 (s, 3H), 2.66-2.48 (m, 1H), 2.38-2.28 (m,3H), 1.79-1.58 (m, 8H); m/z 338.2 (M+H⁺).

Example 31 N-(4-Chlorophenyl)-4-phenoxypiperidine-1-carboxamide

31A. N-(4-Chlorophenyl)-4-oxopiperidine-1-carboxamide

Piperidin-4-one hydrochloride (1.37 g, 10.1 mmol) was dissolved inCH₂Cl₂, washed with 1 M NaOH (60 mL), dried over anhydrous MgSO₄,filtered, and concentrated under reduced pressure to provide the freebase as a clear, colorless oil. The piperidin-4-one was diluted withCH₂Cl₂ (6 mL), and the solution was cooled to 0° C. 4-Chlorophenylisocyanate (1.59 g, 10.1 mmol) was added to the solution, and the icebath was immediately removed. After 3 h, the reaction mixture wasdiluted with brine (10 mL) and 1 M NaOH (2 mL) and extracted with CH₂Cl₂(2×30 mL). The combined organic layers were dried over anhydrous MgSO₄,filtered, and concentrated under reduced pressure to afford the desiredintermediate as a white solid. ¹H NMR (400 MHz; CDCl₃): 8.80 (s, 1H),7.50 (d, J=9.0 Hz, 2H), 7.27 (d, J=8.8 Hz, 2H), 3.72 (t, J=6.2 Hz, 4H),2.38 (t, J=6.2 Hz, 4H); m/z 253.1 (M+H⁺).

31B. N-(4-Chlorophenyl)-4-hydroxypiperidine-1-carboxamide

To a solution of N-(4-chlorophenyl)-4-oxopiperidine-1-carboxamide (401mg, 1.58 mmol) in methanol (20 mL) was added NaBH₄ (89 mg, 2.36 mmol) atrt. The reaction was allowed to stir for 14 h before adding 1 M HCl (20mL). The solution was extracted with CH₂Cl₂ (60 mL), dried overanhydrous MgSO₄, filtered, and concentrated under reduced pressure toafford the desired product as an oil. ¹H NMR (400 MHz; CDCl₃): 8.57 (s,1H), 7.46 (d, J=9.0 Hz, 2H), 7.24 (d, J=9.0 Hz, 2H), 4.70 (d, J=4.3 Hz,1H), 3.79 (td, J=4.3, 13.6 Hz, 2H), 3.68-3.58 (m, 1H), 3.02 (ddd, J=3.2,10.0, 13.3 Hz, 2H), 1.75-1.67 (m, 2H), 1.34-1.21 (m, 2H).

Example 31 N-(4-Chlorophenyl)-4-phenoxypiperidine-1-carboxamide

To a solution of N-(4-chlorophenyl)-4-hydroxypiperidine-1-carboxamide(100 mg, 0.39 mmol) and PPh₃ (430 mg, 1.6 mmol) in THF (2 mL) at rt wasadded diethyl azodicarboxylate (DEAD) (0.068 mL, 0.43 mmol) and phenol(41 mg, 0.43 mmol). The solution was allowed to stir at rt for 16 hbefore concentrating under reduced pressure. The crude residue waspurified using silica gel chromatography (30% EtOAc in hexanes) toafford the desired product as a clear, colorless film. ¹H NMR (400 MHz;CDCl₃): δ 7.34-7.22 (m, 6H), 6.99-6.90 (m, 3H), 6.48 (br s, 1H),4.59-4.54 (m, 1H), 3.75-3.67 (m, 2H), 3.52-3.46 (m, 2H), 2.4-1.97 (m,2H), 1.94-1.86 (m, 2H); m/z 331.2 (M+H⁺).

Example 32 N-(4-Chlorophenyl)-4-phenoxycyclohexane-1-carboxamide

32A. N-(4-Chlorophenyl)-4-hydroxycyclohexane-1-carboxamide

4-Hydroxycyclohexane-1-carboxylic acid (2.0 g, 14 mmol), 4-chloroaniline(1.8 g, 14 mmol), and1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (HATU) (6.3, 17 mmol) were added to a 100 mLround bottom flask followed by DMF (46 mL) and diisopropylethylamine(2.5 mL, 28 mmol). The solution was stirred under argon for 16 h. Thereaction solution was diluted with EtOAc (60 mL), washed with 1 N NaOH(50 mL), dried over anhydrous MgSO₄, filtered, and concentrated underreduced pressure. The crude residue was purified using silica gelchromatography (0% to 100% EtOAc in hexanes) to afford the desiredproduct as a white solid. m/z 254.2 (M+H⁺).

Example 32 N-(4-Chlorophenyl)-4-phenoxycyclohexane-1-carboxamide

To a 50 mL round bottom flask was addedN-(4-chlorophenyl)-4-hydroxycyclohexane-1-carboxamide (1.6 g, 6.3 mmol),polymer-bound PPh₃ (3.0 mmol/g PPh₃, 8.4 g, 25 mmol), and phenol (0.894g, 9.5 mmol). The flask was evacuated and backfilled with argon. To theflask was added THF (30 mL), and the mixture was cooled to 0° C. DEAD(1.49 mL, 9.5 mmol) was added dropwise by syringe, and the ice bath wasremoved. The mixture was allowed to warm to rt and stirred for 16 h. Thereaction mixture was diluted with EtOAc (50 mL), filtered through a padof 1:1 CELITE®:silica gel, and concentrated under reduced pressure. Thecrude residue was purified using silica gel chromatography (0% to 18%,then 18% to 30% EtOAc in hexanes) to afford the desired product as awhite solid. ¹H NMR (400 MHz; CDCl₃): δ 7.48 (d, J=8.7 Hz, 2H),7.32-7.27 (m, 4H), 7.15 (br s, 1H), 6.99-6.84 (m, 3H), 4.34-4.14 (m,1H), 2.36-2.19 (m, 3H), 2.08 (d, J=11.7 Hz, 2H), 1.81-1.68 (m, 2H),1.55-1.43 (m, 2H); m/z 330.2 (M+H⁺).

Example 332-(4-Chlorophenyl)-N-((trans)-4-(4-methoxyphenyl)cyclohexyl)acetamide

33A. cis-4-(4-Methoxyphenyl)cyclohexyl methanesulfonate

To a solution of cis-4-(4-methoxyphenyl)-cyclohexanol (Chem. Commun.,48:9376 (2012)) (366 mg, 1.77 mmol) and triethylamine (0.49 mL, 3.55mmol) in tetrahydrofuran (9 mL), at 0° C., was added methanesulfonylchloride (0.21 mL, 2.66 mmol). The mixture was stirred for 1.5 h beforebeing quenched with water, diluted with EtOAc, then washed sequentiallywith dilute HCl, saturated aqueous solution of sodium bicarbonate, andbrine. The organic phases were dried over sodium sulfate thenconcentrated under reduced pressure before the resultant residue waspurified using silica gel chromatography (20% to 50% EtOAc in hexanes)to afford cis-4-(4-methoxyphenyl)cyclohexyl methanesulfonate, as a whitesolid.

33B. (trans)-4-(4-Methoxyphenyl)cyclohexan-1-amine

To a mixture of cis-4-(4-methoxyphenyl)cyclohexyl methanesulfonate (430mg, 1.51 mmol) in DMF (7.5 mL) was added sodium azide (108 mg, 1.66mmol). The mixture was then heated at 70° C. for 4 h. The mixture wascooled to rt, quenched with water, then diluted with EtOAc. The organicphase was washed several times with water, then brine, before beingconcentrated under reduced pressure to ˜10 mL. To this mixture was addedwet 10% Pd/C (70 mg, 10% w/w) and the reaction vessel was placed under ahydrogen atmosphere, at rt, for 16 h. The mixture was filtered and thefiltrate was concentrated under reduced pressure to afford a residuewhich was purified using silica gel chromatography (10% to 20% methanolin dichloromethane) to furnish the desired product,(trans)-4-(4-methoxyphenyl)cyclohexan-1-amine, as an off-white solid.

Example 332-(4-Chlorophenyl)-N-((trans)-4-(4-methoxyphenyl)cyclohexyl)acetamide

A solution containing 4-chlorophenylacetic acid (109 mg, 0.64 mmol) and1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (266 mg, 0.70 mmol) in DMF (6 mL) was stirredat rt for 10 minutes before(trans)-4-(4-methoxyphenyl)cyclohexan-1-amine (131 mg, 0.64 mmol) wasadded. After stirring for 20 minutes, N,N-diisopropylethylamine (0.33mL, 1.92 mmol) was added, and the mixture stirred for an additional 1 h.The flask contents were then poured into brine (30 mL) and filtered. Thefiltrate was concentrated under reduced pressure to afford a residuewhich was purified using silica gel chromatography (5% MeOH in CH₂Cl₂)to yield the desired2-(4-chlorophenyl)-N-((trans)-4-(4-methoxyphenyl)cyclohexyl)acetamide asa white solid. ¹H NMR (400 MHz; CDCl₃): δ 7.33 (d, J=8.4 Hz, 2H), 7.21(d, J=8.1 Hz, 2H), 7.09 (d, J=8.7 Hz, 2H), 6.83 (d, J=9 Hz, 2H), 5.18(d, J=8.4 Hz, 1H), 3.85-3.78 (m, 4H), 3.52 (s, 2H), 2.43-2.34 (m, 1H),2.05-2.00 (m, 2H), 1.90-1.85 (m, 2H), 1.51-1.04 (m, 4H) ppm. m/z 358.2(M+H)⁺.

Example 344-Fluoro-N-(1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-yl)aniline,HCl

34A. Ethyl 2-(4-(4-hydroxyphenyl)cyclohexylidene)acetate

To an oven-dried flask (Flask #1) was added NaH (60% dispersion in oil,11.8 g, 295 mmol) and 120 mL of THF, before the mixture was cooled to 0°C. To this mixture was added dropwise, over 1 hour, a mixture oftriethylphosphonoacetate (46.9 mL, 236 mmol) in 250 mL of THF. After theaddition was complete, the mixture was stirred for 1 h at rt.

To a separate flask (Flask #2), containing a 0° C. mixture of NaH (60%dispersion in oil, 8.67 g, 216 mmol) in 100 mL THF was carefully added,over 45 minutes, a solution of 37.47 g (196.9 mmol) of4-(4-hydroxyphenyl)cyclohexanone in 250 mL THF. After addition wascomplete, the mixture was stirred at rt for 2 h until the mixture becamea clear solution. Once this solution was clear, Flask #1 was cooled to0° C. and the contents of Flask #2 are added via cannulation. After thisaddition was complete, the mixture was warmed to rt and stirred for 2 h.The mixture was then quenched by careful addition of ice and water (1 L)and subsequently extracted with EtOAc (3×500 mL). The combined organiclayers were then washed with brine (1 L), dried over sodium sulfate,filtered, and concentrated under reduced pressure to provide ethyl2-(4-(4-hydroxyphenyl)cyclohexylidene)acetate in 97% yield as a whitesolid.

34B. Ethyl 2-(4-(4-hydroxyphenyl)cyclohexyl)acetate

To a solution ethyl 2-(4-(4-hydroxyphenyl)cyclohexylidene)acetate (9.74g, 35.8 mmol) in EtOAc was added Pd/C (0.974 g, 10 wt. %). The solutionwas sparged with a balloon of H₂ (g) and stirred under an atmosphere ofhydrogen for 2 days. The mixture was then filtered through a pad ofCELITE®, which was thoroughly rinsed with EtOAc. The combined filtratewas then concentrated under reduced pressure to afford ethyl2-(4-(4-hydroxyphenyl)cyclohexyl)acetate as a white crystalline solid inquantitative yield as a mixture of diastereomers.

34C. Ethyl 2-(4-(4-methoxyphenyl)cyclohexyl)acetate

To a solution of ethyl 2-(4-(4-hydroxyphenyl)cyclohexyl)acetate (34B,34.1 g, 130 mmol) in DMF (300 mL) was added Cs₂CO₃ (65.0 g, 200 mmol)followed by iodomethane (21.3 g, 150 mmol). The resulting suspension wasstirred at rt for 16 h. The mixture was concentrated under reducedpressure and the residue was partitioned between EtOAc (150 ml) andwater (200 mL). The layers were separated and the aqueous layer wasextracted with EtOAc (3×150 mL). These combined organic extracts werecombined with the original organic layer and were dried over anhydrousMgSO₄, filtered, and concentrated under reduced pressure. The residuewas purified employing silica gel chromatography (0% to 30% EtOAc inhexanes) to afford ethyl 2-(4-(4-methoxyphenyl)cyclohexyl)acetate as aclear oil.

34D. 2-(4-(4-Methoxyphenyl)cyclohexyl)ethan-1-ol

Ethyl 2-(4-(4-methoxyphenyl)cyclohexyl)acetate (34C, 1.45 g, 5.25 mmol)was dissolved in THF (26 mL) and cooled to 0° C. LiAlH₄ (596 mg, 15.7mmol) was then added portionwise, and the mixture was warmed to rt over2 h. The mixture was quenched with water, then 2N aqueous HCl, beforebeing extracted with EtOAc. The combined organic phases were washed withbrine, dried over sodium sulfate, and concentrated under reducedpressure to furnish 2-(4-(4-methoxyphenyl)cyclohexyl)ethan-1-ol (1.17 g,95%).

34E. 2-(4-(4-Methoxyphenyl)cyclohexyl)acetaldehyde

To a mixture of 2-(4-(4-methoxyphenyl)cyclohexyl)ethan-1-ol (34D, 1.17g, 5.00 mmol) in dichloromethane (50 mL) was added sodium bicarbonate(1.26 g, 15.0 mmol), followed by Dess-Martin periodinane (3.19 g, 7.5mmol) at 0° C. The resultant mixture was stirred at rt for 16 h, beforebeing diluted with dichloromethane then washed with water. The organiclayer was concentrated under reduced pressure to afford a residue whichwas adsorbed onto silica gel and purified using silica gelchromatography (20% EtOAc in hexanes) to furnish2-(4-(4-methoxyphenyl)cyclohexyl)acetaldehyde (609 mg, 52%) as acolorless oil.

34F. 1,1,1-Trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-ol

A solution of 2-(4-(4-methoxyphenyl)cyclohexyl)acetaldehyde (34E, 609mg, 2.62 mmol) and TMS-CF₃ (0.58 mL, 3.93 mmol) in THF (6 mL) wastreated with a 1M solution of TBAF in THF (15.72 mL, 15.72 mmol) at 0°C. The mixture was allowed to warm to rt for 16 h, then quenched withaqueous 2N HCl (3 mL) over 30 minutes. The mixture was partitionedbetween EtOAc and water and the layers were separated. The organiclayers were washed with brine, dried over sodium sulfate andconcentrated under reduced pressure to afford a residue which waspurified using silica gel chromatography (15% EtOAc in hexanes) todeliver 1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-ol(565 mg, 71%) as a colorless oil.

34G. 1,1,1-Trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-one

To a solution of1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-ol (34F, 565mg, 1.89 mmol) in CH₂Cl₂ (19 mL) was added sodium bicarbonate (476 mg,5.67) followed by Dess-Martin periodinane (1.04 g, 2.46 mmol) at 0° C.The mixture was stirred at rt for 16 h, then diluted with CH₂Cl₂ andwashed with water. The mixture was concentrated under reduced pressureand the residue was purified using silica gel chromatography (10% EtOAcin hexanes) to afford1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-one (356 mg,63%) as a colorless oil which crystallized upon standing.

34H.N-(4-Chlorophenyl)-1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-imineand(Z)-4-chloro-N-(3,3,3-trifluoro-1-(4-(4-methoxyphenyl)cyclohexyl)prop-1-en-2-yl)aniline

A mixture of1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-one (34G, 178mg, 0.59 mmol), 4-fluoroaniline (0.14 mL, 1.19 mmol) and p-TsOH (5 mg,0.03 mmol), dissolved in toluene (5 mL), was heated to reflux utilizinga Dean-Stark trap for 16 h. The mixture was concentrated under reducedpressure to afford a residue which was purified by column chromatographyon neutral alumina (7% EtOAc/hexanes) to give a mixture of imineN-(4-chlorophenyl)-1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-imineand(Z)-4-chloro-N-(3,3,3-trifluoro-1-(4-(4-methoxyphenyl)cyclohexyl)prop-1-en-2-yl)anilineas a viscous, colorless oil.

Example 344-Fluoro-N-(1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-yl)aniline

To a mixture of the product of 34H(N-(4-chlorophenyl)-1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-imineand(Z)-4-chloro-N-(3,3,3-trifluoro-1-(4-(4-methoxyphenyl)cyclohexyl)prop-1-en-2-yl)aniline)(34H, 160 mg, 0.41 mmol) in MeOH (10 mL), at rt, was added sodiumborohydride (46 mg, 1.2 mmol). The resultant mixture was stirred at rtfor 1.5 h before being quenched with satd. aq. ammonium chloride thenextracted with dichloromethane. The combined organic phases were driedover sodium sulfate and concentrated under reduced pressure. Theresultant residue was purified by preparative HPLC (Varian ProStar usingHamilton C18 PRP-1 column (15×250 mm) with flow rate of 20 mL/min,Mobile Phase A: 0.5% formic acid in water; Mobile Phase B: 0.5% formicacid in acetonitrile; 0% to 100% B gradient elution during 30 minutes)to give a residue. The residue was diluted with 2M HCl in diethyl etherand concentrated under reduced pressure to give the HCl salt of thedesired compound as a racemic mixture of cis/trans isomers. ¹H NMR (300MHz; CDCl₃): δ 7.16-7.08 (m, 2H), 6.95-6.81 (m, 4H), 6.65-6.59 (m, 2H),3.93-3.85 (m, 1H), 3.85-3.71 (m, 3H), 3.41 (d, J=9.0 Hz, 1H), 2.55-2.50(m, 1H), 2.41 (tt, J=12.3, 3.0 Hz, 1H), 2.17-2.08 (m, 1H), 1.99-1.00 (m,9H) ppm. m/z 396.15 (M+H)⁺.

Example 354-Chloro-N-(1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-yl)anilinehydrogenchloride

Prepared utilizing the procedures used to afford4-fluoro-N-(1,1,1-trifluoro-3-(4-(4-methoxyphenyl)cyclohexyl)propan-2-yl)anilinereplacing 4-fluoroaniline with 4-chloroaniline. MS(ES): m/z=412.10[M+H]⁺. t_(R)=2.94 min (Method M).

Example 36 2-(4-Cyanophenyl)-N-((trans)-4-phenylcyclohexyl)acetamide

The following compound was made in the manner of General Procedure A,employing trans-4-phenyl-cyclohexanamine (PCT Publication No. WO2001/092204) (138 mg, 0.79 mmol), 2-(4-cyanophenyl)acetic acid (138 mg,0.86 mmol),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (360 mg, 0.95 mmol), and DMF (4 mL). Theresidue was purified to by preparative TLC (33% EtOAc in hexanes),followed by preparative HPLC (Varian ProStar using Hamilton C18 PRP-1column (15×250 mm) with flow rate of 20 mL/min, Mobile Phase A: 0.5%formic acid in water; Mobile Phase B: 0.5% formic acid in acetonitrile;0% to 100% B gradient elution during 30 minutes) to give the desiredproduct. ¹H NMR (300 MHz; CDCl₃): δ 7.64 (dd, J=8.4, 1.8 Hz, 2H), 7.40(dd, J=8.4, 1.8 Hz, 2H), 7.32-7.24 (m, 2H), 7.21-7.16 (m, 3H), 5.31 (d,J=7.5 Hz, 1H), 3.89-3.79 (m, 1H), 3.60 (s, 2H), 2.45 (tt, J=16.0, 3.6Hz, 1H), 2.10-1.90 (m, 4H), 1.66-1.56 (m, 2H), 1.30-1.16 (m, 2H) ppm.m/z 319 (M+H)⁺.

Example 37 2-(4-Chlorophenyl)-N-((trans)-4-phenylcyclohexyl)propanamide

The following compound was made in the manner of General Procedure A,employing trans-4-phenyl-cyclohexanamine (PCT Publication No. WO2001/092204) (138 mg, 0.79 mmol), 2-(4-chlorophenyl)propanoic acid (158mg, 0.86 mmol),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (360 mg, 0.95 mmol), and DMF (4 mL). Theresidue was purified to by preparative TLC (33% EtOAc in hexanes) togive a residue. The residue was further purified by preparative HPLC(Varian ProStar using Hamilton C18 PRP-1 column (15×250 mm) with flowrate of 20 mL/min, Mobile Phase A: 0.5% formic acid in water; MobilePhase B: 0.5% formic acid in acetonitrile; 0% to 100% B gradient elutionduring 30 minutes) to give the desired product as a racemic mixture. ¹HNMR (300 MHz; CDCl₃): δ 7.34-7.14 (m, 9H), 5.16 (d, J=7.8 Hz, 1H),3.86-3.75 (m, 1H), 3.48 (q, J=7.2 Hz, 1H), 2.42 (tt, J=12.3, 3.3 Hz,1H), 2.09-1.84 (m, 4H), 1.68-1.52 (m, 2H), 1.49 (d, J=7.2 Hz, 3H),1.26-1.06 (m, 2H) ppm. m/z 342 (M+H)⁺.

Example 382-(4-Chlorophenyl)-N-((trans)-4-(quinolin-4-yl)cyclohexyl)acetamide

38A. 4-(1,4-Dioxaspiro[4.5]dec-7-en-8-yl)quinoline

1,4-Dioxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate (Bioorg. Med.Chem. Lett., 24:5377 (2014)) (6.9 g, 23.9 mmol) was placed in a 500 mLround bottomed flask, followed by quinolone-4-boronic acid (4.55 g, 26.3mmol), Pd(PPh₃)₄ (1.39 g, 1.2 mmol, 5 mol %), KBr (2.85 g, 23.94 mmol)and sodium carbonate (6.34 g, 59.85 mmol). The flask was evacuated andbackfilled with N₂ three times, before degassed dioxane (100 mL) andwater (10 mL) were added to the solids and the mixture was stirred andheated to 90° C. under N₂ atmosphere. After 16 h, the mixture was cooledto rt, and SiO₂ was added. The mixture was concentrated under reducedpressure and the residue was purified by silica gel columnchromatography (0% to 100% EtOAc in hexanes) to give4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)quinoline (3.2 g, 50%).

38B. 4-(1,4-Dioxaspiro[4.5]decan-8-yl)quinolone

A mixture 4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)quinolone (3.2 g, 12.0mmol), NaHCO₃ (500 mg, 6.0 mmol), and MeOH (70 mL) was purged with N₂(g), before 20 wt. % of Pd/C (dry activated, 10 wt. %) was added to themixture. H₂ (g) was bubbled through the solution until completedisappearance of the starting material. The mixture was purged with N₂(g), filtered through CELITE®, and the filtrate was concentrated underreduced pressure. The residue was purified by flash chromatography togive 4-(1,4-dioxaspiro[4.5]decan-8-yl)quinolone (2.9 g, 90%).

38C. 4-(Quinolin-4-yl)cyclohexan-1-one

To a mixture of 4-(1,4-dioxaspiro[4.5]decan-8-yl)quinolone (3.0 g, 11mmol) in acetone (30 mL) was added 3M aqueous HCl (30 mL). After themixture was stirred at rt for 24 h, it was concentrated under reducedpressure and NaHCO₃ (sat. aqueous solution) was added to adjust the pHabove 8.0. The mixture was then extracted with EtOAc. The combinedorganic layers were dried with Na₂SO₄, and concentrated under reducedpressure to furnish 4-(quinolin-4-yl)cyclohexan-1-one as a yellow oil(2.3 g, 90%), which solidified upon standing.

38D. 4-(Quinolin-4-yl)cyclohexan-1-amine

The desired amine was made through reductive amination of4-(quinolin-4-yl)cyclohexan-1-one with ammonium acetate and sodiumcyanoborohydride to give 4-(quinolin-4-yl)cyclohexan-1-amine.

38E. 2-(4-Chlorophenyl)-N-((trans)-4-(quinolin-4-yl)cyclohexyl)acetamide

General Procedure A was employed using4-(quinolin-4-yl)cyclohexan-1-amine, and 2-(4-chlorophenyl)acetic acid.The residue was purified using preparative HPLC (Varian ProStar usingHamilton C18 PRP-1 column (15×250 mm) with flow rate of 20 mL/min,Mobile Phase A: 0.5% formic acid in water; Mobile Phase B: 0.5% formicacid in acetonitrile; 0% to 100% B gradient elution during 30 minutes)to give the desired compound as a white powder. ¹H NMR (400 MHz; CD₃OD):δ 8.77-8.74 (m, 1H), 8.26-8.21 (m, 1H), 8.05-8.01 (m, 1H), 7.79-7.73 (m,1H), 7.68-7.63 (m, 1H), 7.49-7.45 (m, 1H), 7.34-7.27 (m, 4H), 3.82-3.77(m, 1H), 3.50-3.42 (m, 3H), 2.13-2.03 (m, 4H), 1.82-1.70 (m, 2H),1.65-1.60 (m, 2H) ppm. m/z 379 (M+H)⁺.

Example 40N-((R)-1-((1s,4S)-4-(6-Fluoroquinolin-4-yl)cyclohexyl)ethyl)-3-methylbenzenesulfonamide

Preparation 40A:

To a stirred solution of 1,4-dioxaspiro[4.5]decan-8-one (300 g, 1920.86mmol, 1.0 eq) and phenyltrifluoromethanesulfonimide (823.47 g, 2305.03mmol, 1.2 eq) in MTBE (7.5 L) under N₂ at −78° C. was added 2.0 M NaHMDSin THF (1152.2 mL, 2305.03 mmol, 1.2 eq) over 70 minutes, and themixture was stirred for an additional 60 minutes. The reaction mixturewas warmed to room temperature and stirred overnight until TLC showedcomplete consumption of the starting material. The mixture was quenchedwith aqueous KHSO₄ (100 ml), filtrated to remove the solid andconcentrated the filtrate completely. To the residue was added 3 L MTBE,then washed with 5% NaOH (1.5 L×3). The organic phase was concentratedto obtain 567 g crude Preparation 40A (light yellow oil, yield 102%).The crude can be used directly in next step without furtherpurification.

Preparation 40A: ¹H NMR (400 MHz, CDCl₃): δ (ppm) 5.65 (t, J=4.0 Hz,1H), 3.98 (d, J=1.5 Hz, 4H), 2.53 (s, 2H), 2.40 (s, 2H), 1.90 (t, J=6.6Hz, 2H)

Preparation 40B:

A mixture of crude Preparation 40A (600 g, 2.08 mol, 1 eq), B₂Pin₂(687.1 g, 2.71 mol, 1.3 eq), KOAc (613 g, 6.24 mol, 3 eq), NaBr (86 g0.833 mol, 0.4 eq) and Pd(dppf)Cl₂ (76 g, 0.1 mol, 0.05 eq) in dioxane(6.5 L) was heated to reflux overnight. Once the reaction was complete,the mixture was concentrated and purified by FCC (2%→10%→20% EtOAc/PE)to give Preparation 40B (369 g, 66%).

Preparation 40B: LC-MS: 267.1 (M+1)+, ¹H NMR (400 MHz, CDCl₃) δ 6.46 (s,1H), 3.98 (s, 4H), 2.37-2.35 (m, 4H), 1.74-1.60 (t, 2H), 1.24 (s, 12H).

Preparation 40C:

A mixture of Preparation 40B (368 g, 1.38 mol, 1.3 eq),4-Chloro-6-fluoroquinoline (195 g, 1.07 mol, 1 eq), K₂CO₃ (445 g, 3.22mol, 3 eq) and Pd(PPh₃)₄ (25 g, 22 mmol, 0.02 eq) in dioxane-water (3 L,4:1) was heated to reflux overnight. The solution was then concentratedand extracted with EtOAc. Purification by FCC (38% EtOAc/petroleumether) gave Preparation 40C (236 g, 77%).

Preparation 40C: LC-MS: 286.1 (M+1)+, ¹H NMR (400 MHz, CDCl₃) δ8.80-8.29 (d, 1H), 8.11-8.07 (q, 1H), 7.63-7.61 (q, 1H), 7.47-7.46 (q,1H), 7.26-7.22 (m, 1H), 5.75-5.74 (m, 1H), 4.08-4.05 (m, 4H), 2.63-2.59(m, 2H), 2.59-2.53 (m, 2H), 2.0-1.97 (m, 2H).

Preparation 40D:

To Preparation 40C (125 g, 0.44 mol) in IPA (2 L) at 55° C. was added10% Pd/C and the mixture was stirred under an atmosphere of H₂overnight. The mixture was filtered and concentrated to give crudePreparation 40D (130 g), which was used directly in the next step.

Preparation 40E:

Preparation 40D (100 g, 0.348 mol) was treated with 4 N HCl (300 mL) inacetone (1200 mL) at 45° C. overnight. The mixture was monitored by TLC.Then the solution was concentrated in vacuo. The residue was adjusted topH 9 with 6 N NaOH. and the mixture was partitioned between ethylacetate and water. The organic layer was washed with brine, dried overanhydrous Na₂SO₄, filtered and concentrated to give light yellow solid,which was then purified by silica gel column using hexanes and ethylacetate (from 20 percent ethyl acetate to 70% ethyl acetate) to affordPreparation 40E as a white solid, (47 g+20 g mixture, yield >55%).Preparation 40E: LC-MS: 244.0 (M+1)+, ¹H NMR (400 MHz, CDCl₃) δ 8.84 (d,J=4.6 Hz, 1H), 8.16 (dd, J=9.3, 5.7 Hz, 1H), 7.72 (dd, J=10.3, 2.8 Hz,1H), 7.52 (ddd, J=9.2, 7.8, 2.7 Hz, 1H), 7.29 (d, J=4.6 Hz, 1H), 3.69(ddd, J=12.1, 9.0, 3.3 Hz, 1H), 2.77-2.54 (m, 4H), 2.37 (ddd, J=13.4,5.9, 3.0 Hz, 2H), 2.04 (qd, J=12.6, 5.3 Hz, 2H).

Preparation 40F:

Preparation 40E (57.8 g, 237.8 mmol) was dissolved in EtOH (240 mL) andcooled to 0° C. NaBH₄ (9.94 g, 261.6 mmol) was added portionwisemaintaining the temperature within a range of 0-10° C. (exothermicreaction). The resulting suspension was stirred for 20 minutes. An LC/MSof an aliquot of the reaction mixture indicated consumption of ketone(m/z (M+H)+=244). The reaction was quenched at 0° C. by the slowaddition of acetone (58 mL) over 15 minutes (exotherm). The reaction waspoured slowly onto 500 mL of saturated aqueous ammonium chloride and 500g of ice. The resulting aqueous solution was extracted with EtOAc (3×300mL) and the combined organic fractions were washed with saturatedaqueous ammonium chloride (250 mL) and saturated aqueous sodium chloride(250 mL). The organic portion was dried over anhydrous sodium sulfateand concentrated under reduced pressure. Sufficient silica to adsorb theoil was added and diluted with 10% MeOH in CH₂Cl₂. A similar quantity ofsilica was used as a silica plug to purify the material. The silica plugwas washed with 10% MeOH in CH₂Cl₂ until UV-active material no longercould be detected by TLC (7:3 EtOAc/Hexanes, R_(f)=0.4). The filtratewas concentrated then suspended in 500 mL of toluene and concentratedagain. Crude Preparation 40F was isolated as a yellow solid (58.2 g)that was used in the subsequent step without further purification.

Preparation 40G:

To Preparation 40F (58.2 g, 237.8 mmol) was added MeCN (125 mL) andpyridine (38.7 mL, 480 mmol) and the reaction mixture was cooled to 5°C. using an ice/water bath. Methanesulfonyl chloride (26.0 mL, 336 mmol)was added dropwise at 5° C. (exothermic reaction), the reaction mixturestirred for 1 hr at 5° C. and then brought up to room temperature andstirred for an additional 16 h during which time a white precipitateformed. The heterogeneous mixture was quenched by the addition ofsaturated aqueous ammonium chloride (200 mL) and extracted with CH₂Cl₂(3×300 mL). The combined organic fractions were dried over anhydroussodium sulfate and concentrated under reduced pressure. Excess pyridinewas removed by azeotroping from toluene (3×300 mL). The crude materialwas recrystallized from H₂O/MeOH as follows: 1 mL/mmol of H₂O was addedand the slurry was heated to 120° C. in an oil bath. MeOH was addeduntil the solids went into solution (˜0.5 L). After cooling whitecrystals were collected by filtration to give Preparation 40G (58.6g, >20:1 dr, 76% over two steps). m/z (M+H)+=324.1. H-NMR (400 MHz;CDCl₃): δ 8.82 (dd, J=4.6, 0.2 Hz, 1H), 8.15-8.11 (m, 1H), 7.64-7.61 (m,1H), 7.52-7.46 (m, 1H), 7.25 (s, 1H), 4.78 (tt, J=10.9, 5.2 Hz, 1H),3.24-3.16 (m, 1H), 3.07 (d, J=1.0 Hz, 3H), 2.42-2.38 (m, 2H), 2.16-2.12(m, 2H), 1.93-1.66 (m, 4H).

Preparation 40H:

Di-tert-butyl malonate (33.5 mL, 150 mmol) was added dropwise to astirred suspension of NaH (6.0 g, 60% suspension in oil, 150 mmol) in1,2-dimethoxyethane (100 mL) under Ar, cooled in a water-ice bath. Afterstirring for 10 min, Preparation 40G (16.2 g, 50 mmol) was added and thereaction was heated at 85° C. for 20 h. After this time, acetic acid(100 mL) was added, the reaction flask was fitted with a distillationhead and the temperature was raised to 130° C. 1,2-dimethoxyethane wasdistilled off under atmospheric pressure until the distillate was acidic(˜100 mL). The distillation head was removed, a reflux condenser wasattached, water (20 mL) was added and the reaction heated at 130° C. for12 h. The reaction was concentrated under reduced pressure and pouredonto 200 g of ice and 100 mL of saturated aqueous NaOAc. Preparation 40Hwas isolated as a white solid by filtration and further dried byrefluxing with toluene in a Dean-Stark apparatus (11.0 g, 76%). m/z(M+H)⁺=288.2. ¹H-NMR (400 MHz; DMSO-d₆): δ 12.05 (bs, 1H), 8.79 (d,J=4.5 Hz, 1H), 8.06 (dd, J=9.2, 5.8 Hz, 1H), 7.94 (dd, J=11.0, 2.8 Hz,1H), 7.66-7.61 (m, 1H), 7.50 (d, J=4.6 Hz, 1H), 2.41 (d, J=7.6 Hz, 2H),2.28-2.23 (m, 1H), 1.87-1.78 (m, 2H), 1.73-1.64 (m, 6H).

Preparation 40I:

To a solution of Preparation 40H (1.4 g, 4.8 mmol) in THF (15 mL) wasadded NEt₃ (1.3 mL, 9.6 mmol). The reaction mixture was cooled to 0° C.and trimethylacetyl chloride (0.713 mL, 5.8 mmol) was added dropwise andthe resulting solution stirred for 30 min at 0° C. In a separate flask,(R)-4-phenyloxazolidin-2-one (3, 1.01 g, 6.24 mmol) in THF (45 mL) at 0°C. was treated with 1 M LiHMDS solution in THF (dropwise addition of6.24 mL, 6.24 mmol) and stirred at 0° C. The lithiate was added viacannula to the first flask. The reaction mixture was allowed to warm tort and was stirred for 3 hours. LC/MS indicated the complete consumptionof the starting carboxylic acid and formation of the desired imide. Thereaction mixture was poured onto saturated aqueous ammonium chloride (50mL) and the layers were separated. The aqueous layer was extracted withEtOAc (3×50 mL). The combined organic extracts were dried over anhydroussodium sulfate and chromatographed on silica using EtOAc/Hexanes 0 to100% gradient to give Preparation 40I as a white foam in 83% yield. m/z(M+H)⁺=433.3. ¹H-NMR (400 MHz; CDCl₃): δ 8.80 (d, J=4.5 Hz, 1H), 8.11(dd, J=9.1, 5.7 Hz, 1H), 7.63 (dd, J=10.5, 2.5 Hz, 1H), 7.48-7.43 (m,1H), 7.40-7.30 (m, 6H), 5.47-5.44 (m, 1H), 4.71 (t, J=8.9 Hz, 1H),4.31-4.28 (m, 1H), 3.20-3.11 (m, 3H), 2.49-2.46 (m, 1H), 1.82-1.67 (m,6H).

Preparation 40J:

A solution of Preparation 40I (21.6 g, 50 mmol) in anhydrous THF (200mL) was cooled to −40° C. (using acetonitrile/dry ice bath, someprecipitation occurs) and 2 M NaHMDS solution in THF (30 mL, 60 mmol)was added over 5 min (a 5-8° C. rise in temperature was observed). Theresulting yellow reaction mixture was stirred for 10 min, becamehomogeneous, and MeI (10.6 g, 75 mmol) was added dropwise over 2 min (a10° C. rise in temperature was observed). The reaction mixture wasstirred for 1 h at −40° C. and LC/MS indicated the complete consumptionof the starting material and formation of the desired methyl imide. Thereaction mixture was rapidly diluted with saturated aqueous ammoniumchloride solution (400 mL) and the biphasic mixture was stirred for 15min. ^(i)PrOAc (100 mL) was added, the layers were separated, and theaqueous layer was extracted with ^(i)PrOAc (3×50 mL). The combinedorganic extracts were dried over anhydrous magnesium sulfate filtered,and concentrated. The resulting residue was recrystallized by dissolvingin 400 mL hot acetone and adding H₂O until a milky solution formedfollowed to re-dissolving with heating (˜3:1 acetone/H₂O). Preparation40J was obtained as white needles (15.04 g, 2 crops, 68%). m/z(M+H)⁺=447.3. ¹H-NMR (400 MHz; CDCl₃): δ 8.81 (d, J=4.6 Hz, 1H), 8.10(dd, J=9.2, 5.7 Hz, 1H), 7.65 (dd, J=10.6, 2.7 Hz, 1H), 7.47-7.42 (m,1H), 7.41-7.29 (m, 6H), 5.47 (dd, J=8.8, 3.8 Hz, 1H), 4.69 (t, J=8.9 Hz,1H), 4.38-4.30 (m, 1H), 4.26 (dd, J=8.9, 3.9 Hz, 1H), 3.26-3.21 (m, 1H),2.18-2.15 (m, 1H), 1.93-1.64 (m, 8H), 1.09 (d, J=6.9 Hz, 3H).

Preparation 40K:

To a solution of Preparation 40J (82.0 g, 183.6 mmol) in THF (610 mL) at0° C. was added aqueous H₂O₂ (35 wt %, 82 mL) and LiOH (7.04 g, 293.8mmol) in H₂O (189 mL). The resulting reaction mixture was allowed toslowly warm to rt and stirred overnight. The reaction was cooled to 0°C. and saturated aqueous sodium bisulfite solution (250 mL) was added.After stirring for 30 min, the THF was removed under reduced pressure.Acetic acid (34 mL) was added followed by EtOAc (300 mL). The layerswere separated, and the aqueous layer was extracted with EtOAc (3×500mL). The combined organic extracts were dried over anhydrous Na₂SO₄,filtered, and concentrated under reduced pressure. The brown crudereaction mixture was suspended in MeCN (400 mL) and the suspension wasbrought to reflux with vigorous stirring. After cooling to rt, thesolids were collected by filtration washing with additional MeCN.Preparation 40K was obtained as a white solid (45.4 g, 82%). m/z(M+H)⁺=302.2. ¹H-NMR (400 MHz; DMSO-d6): δ 12.10 (s, 1H), 8.79 (d, J=4.5Hz, 1H), 8.07 (dd, J=9.2, 5.9 Hz, 1H), 7.97-7.94 (m, 1H), 7.67-7.62 (m,1H), 7.49 (d, J=4.5 Hz, 1H), 3.41-3.36 (m, 1H), 2.73-2.65 (m, 1H),1.83-1.61 (m, 9H), 1.08 (d, J=6.8 Hz, 3H). Chiral HPLC, >99% ee(ChiralPak IC-3, 3 μM, 4.6×250 mm, 15 min isocratic 70% heptane 30%i-PrOH with 230 nm detection) at a flow rate of 0.75 mL/min the desiredenantiomer had a retention time of 8.6 min with the undesired enantiomereluting at 9.5 min.

Preparation 40L:

Preparation 40K (2 g, 6.64 mmol) was taken up in toluene (22.12 ml) anddiphenyl phosphorazidate (2.009 g, 7.30 mmol) and triethylamine (1.110ml, 7.96 mmol) were added. Vial sealed and heated to 70° C. After 2hours, the reaction was cooled to room temperature and concentratedunder reduced pressure. Crude residue was taken up in 40 mL THF and 40mL of water and lithium hydroxide (1.589 g, 66.4 mmol) was added. Thereaction was stirred at room temperature for 1 hour. The reaction wasacidified with 1N HCl (white precipitate forms) and extracted withEtOAc. The aqueous portion was then basified with 1N NaOH (precipitateforms) and extracted with EtOAc 5 times. Basic extracts wereconcentrated in vacuo to give 40L (1.68 g, 6.17 mmol, 93% yield). LC-MSAnal. Calc'd for C₁₇H₂₁FN₂ 272.17. found [M+H] 273.1 T_(r)=0.50 min(Method A). ¹H NMR (400 Mhz, chloroform-d) δ: 8.80 (d, J=4.6 Hz, 1H),8.11 (dd, J=9.3, 5.7 Hz, 1H), 7.67 (dd, J=10.6, 2.8 Hz, 1H), 7.46 (ddd,J=9.2, 8.0, 2.8 Hz, 1H), 7.32 (d, J=4.5 Hz, 1H), 3.27-3.37 (m, 1H), 3.13(dq, J=9.3, 6.3 Hz, 1H), 2.01-2.10 (m, 1H), 1.67-1.92 (m, 6H), 1.37-1.55(m, 4H), 1.15 (d, J=6.4 Hz, 3H).

Example 40N-((R)-1-((1s,4S)-4-(6-Fluoroquinolin-4-yl)cyclohexyl)ethyl)-3-methylbenzenesulfonamide

Preparation 40L (20 mg, 0.073 mmol) was dissolved DCM (0.2 mL) and addedto a vial containing phenyl sulfonyl chloride (26 mg, 0.147 mmol) andDCM (0.2 mL) followed directly after with the addition of DIPEA (64.1μl, 0.367 mmol). The reaction was stirred at room temperature overnight.After overnight, the reaction was concentrated in vacuo, taken up in 2mL DMF, filtered, and purified via HPLC to give Example 40 (12.2 mg,0.28 mmol, 39%) LC-MS Anal. Calc'd for C₂₄H₂₇FN₂O₂S 426.18. found [M+H]427.3 T_(r)=2.065 min (Method B). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.82 (d,J=4.5 Hz, 1H), 8.08 (dd, J=9.2, 5.8 Hz, 1H), 7.90 (dd, J=10.9, 2.5 Hz,1H), 7.60-7.72 (m, 3H), 7.50 (d, J=8.5 Hz, 1H), 7.45 (t, J=7.7 Hz, 1H),7.38 (d, J=7.6 Hz, 1H), 7.32 (d, J=4.5 Hz, 1H), 3.29 (t, J=10.5 Hz, 1H),2.35 (s, 3H), 1.64-1.83 (m, 3H), 1.41-1.64 (m, 4H), 1.23-1.41 (m, 2H),0.92 (d, J=6.4 Hz, 3H).

Examples 41 to 46

Examples 41 to 46 were prepared from Preparation 40L following theprocedure for Example 40 using the corresponding sulfonyl chlorides.

Tr Ex. No. Name R (min)^((Method B)) [M + H]⁺ 41 N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl) ethyl)-4-methylbenzenesulfonamide

2.065 427.3 42 2-chloro-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl) ethyl)benzenesulfonamide

2.085 447.2 43 4-chloro-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl) ethyl)benzenesulfonamide

2.126 447.2 44 3-chloro-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl) ethyl)benzenesulfonamide

2.124 447.2 45 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-3- methoxybenzenesulfonamide

1.995 443.3 46 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4- methoxybenzenesulfonamide

1.975 443.3

Example 474-Chloro-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

Preparation 40L (4 mg, 0.015 mmol) was taken up in DMF (147 μl) and HOBT(2.92 mg, 0.019 mmol), EDC (3.66 mg, 0.019 mmol), 4-chlorobenzoic acid(4.60 mg, 0.029 mmol) and TEA (10.23 μl, 0.073 mmol) were added andreaction stirred at room temperature overnight. The reaction was thendiluted with 1.8 mL DMF and purified via preparative HPLC to giveExample 47 (2.3 mg, 0.006 mmol, 38%). LC-MS Anal. Calc'd for C₂₄H₂₄ClFN₂410.16. found [M+H] 411.0 T_(r)=2.057 min (Method B). ¹H NMR (500 MHz,DMSO-d₆) δ: 8.82 (d, J=4.5 Hz, 1H), 8.36 (d, J=8.8 Hz, 1H), 8.08 (dd,J=9.2, 5.8 Hz, 1H), 7.95 (dd, J=11.0, 2.7 Hz, 1H), 7.86 (d, J=8.6 Hz,2H), 7.65 (td, J=8.7, 2.7 Hz, 1H), 7.52 (d, J=8.5 Hz, 2H), 7.47 (d,J=4.5 Hz, 1H), 4.42 (d, J=6.6 Hz, 1H), 1.74-1.91 (m, 6H), 1.56-1.73 (m,4H), 1.18 (d, J=6.5 Hz, 3H).

Examples 48 to 69

Examples 48 to 69 were prepared from Preparation 40L following theprocedure for Example 47 using the corresponding benzoic acids.

Ex. Tr No. Name R (min)^((Method B)) [M + H]⁺ 48 N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4- yl)cyclohexyl)ethyl)benzamide

1.875 377.1 49 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-2- methylbenzamide

1.949 391.1 50 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-3-methyl benzamide

2.004 391.3 51 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4- methylbenzamide

2.035 391.3 52 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-2- methoxybenzamide

2.055 407.3 53 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-3- methoxybenzamide

1.955 407.3 54 2-fluoro-N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

1.971 395.1 55 3-fluoro-N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

1.985 395.3 56 2-chloro-N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

1.985 411.3 57 3-chloro-N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

2.115 411.2 59 3,4-dichloro-N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

2.277 445.2 60 4-fluoro-N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

1.955 395.3 61 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-[1,1′-biphenyl]- 3-carboxamide

2.307 453.3 66 3,5-dichloro-N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

2.348 445.2 67 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)picolinamide

1.945 378.1 68 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4- methoxybenzamide

1.914 407.3 69 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-[1,1′-biphenyl]- 2-carboxamide

2.206 453.3

Example 70N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)-[1,1′-biphenyl]-4-carboxamide

Example 70N-((R)-1-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)-[1,1′-biphenyl]-4-carboxamide

Preparation 40L (50 mg, 0.184 mmol) was taken up in DMF (1836 μl) andHOBT (36.5 mg, 0.239 mmol), EDC (45.8 mg, 0.239 mmol),[1,1′-biphenyl]-4-carboxylic acid (54.6 mg, 0.275 mmol) and TEA (128 μl,0.918 mmol) were added and reaction stirred at room temperature for 3hours. Reaction diluted with EtOAc and washed with 5:1 water/aqueoussaturated NaHCO₃ solution. The combined organic extracts were dried withsodium sulfate, filtered and concentrated in vacuo. The crude residuewas purified via silica gel flash column chromatography to give Example70 (63 mg, 0.132 mmol, 72.0% yield). LC-MS Anal. Calc'd for C₃₀H₂₉FN₂O452.23. found [M+H] 453.3 T_(r)=2.297 min (Method B). ¹H NMR (400 MHz,CHLOROFORM-d) δ: 8.82 (d, J=4.5 Hz, 1H), 8.13 (dd, J=9.2, 5.7 Hz, 1H),7.85 (d, J=8.6 Hz, 2H), 7.64-7.70 (m, 3H), 7.58-7.64 (m, J=7.0 Hz, 2H),7.36-7.50 (m, 5H), 5.91 (d, J=9.2 Hz, 1H), 4.58-4.70 (m, 1H), 3.30 (tt,J=10.6, 3.6 Hz, 1H), 1.96-2.15 (m, 3H), 1.80 (br. s., 6H), 1.33 (d,J=6.6 Hz, 3H).

Example 714-Chloro-N-(1-((trans)-4-(quinolin-4-yloxy)cyclohexyl)propyl)benzamide

Intermediate 71A. Ethyl 2-(1,4-dioxaspiro[4.5]decan-8-ylidene)acetate

Triethyl phosphonoacetate (21.79 ml, 109 mmol) was added to a suspensionof sodium hydride (3.84 g, 96 mmol) in THF (64.0 ml) and 0° C. Reactionwas stirred at room temperature for 30 minutes. After 30 minutes, thereaction was recooled to 0° C. and a solution of1,4-dioxaspiro[4.5]decan-8-one (10 g, 64.0 mmol) in 5 mL THF was added.The reaction was then stirred at room temperature for 30 minutes priorto quenching with water. The mixture was extracted with DCM three times.Combined organic extracts were dried with sodium sulfate, filtered, andconcentrated in vacuo. Crude residue was purified via silica gelchromatography to give Intermediate 71A (13.88 g, 61.3 mmol, 96% yield).TLC: product stains as purple spot in anisaldehyde (Rf=0.75 in 1:1Hex/EtOAc). ¹H NMR (400 MHz, chloroform-d) δ: 5.65 (s, 1H), 4.13 (q,J=7.2 Hz, 2H), 3.92-3.99 (m, 4H), 2.94-3.02 (m, 2H), 2.31-2.40 (m, 2H),1.71-1.79 (m, 4H), 1.26 (t, J=7.2 Hz, 3H).

Intermediate 71B. Ethyl 2-(1,4-dioxaspiro[4.5]decan-8-yl)acetate

Intermediate 71A (13.88 g, 61.3 mmol) was taken up in EtOAc (61.3 ml)and was added to a Parr hydrogenation bottle containing wet 10%palladium on carbon (1.306 g, 12.27 mmol) (54% w/w water) under anatmosphere of nitrogen. The reaction bottle was purged and back-filledwith nitrogen three times, and then with hydrogen. After filling thebottle with hydrogen to 50 psi, the bottle was placed in a Parr shakerand shaken. After 4 hours, the reaction was filtered over pressedCELITE® and concentrated in vacuo to give Intermediate 71B (13.78 g,60.4 mmol, 98% yield). LC-MS Anal. Calc'd for C₁₂H₂₀O₄ 228.14. found[M+H] 229.1 T_(r)=0.83 min (Method A). ¹H NMR (400 MHz, chloroform-d) δ:4.11 (q, J=7.2 Hz, 2H), 3.88-3.95 (m, 4H), 2.2.1 (d, J=7.0 Hz, 2H), 1.83(dqd, J=11.0, 7.5, 3.5 Hz, 1H), 1.68-1.78 (m, 4H), 1.50-1.61 (m, 2H),1.27-1.35 (m, 2H), 1.24 (t, J=7.2 Hz, 3H).

Intermediate 71C. Ethyl 2-(1,4-dioxaspiro[4.5]decan-8-yl)butanoate

Diisopropylamine (2.347 ml, 16.63 mmol) taken up in dry THF (15.99 ml)(under N₂ atmosphere) and cooled to −78° C. n-BuLi (6.14 ml, 15.35 mmol)(2.5 M in hexanes) was added over ˜5 minutes at −78° C. After stirringfor 45 minutes, reaction was warmed to room temperature for 10 minutesand returned to −78° C. Then, 1,3-dimethyltetrahydropyrimidin-2(1H)-one(1.541 ml, 12.79 mmol) was added followed by a solution of Intermediate71B (2.92 g, 12.79 mmol) in THF (15.99 ml) (dropwise over ˜5 minutes).After 1 hour, iodoethane (1.125 ml, 14.07 mmol) (neat) was addeddropwise over ˜5 minutes. Reaction stirred another 2 hours at −78° C.before slowly warming to room temperature. The reaction was then stirredover night at room temperature. The reaction was quenched by pouringinto 1:1 water/brine and extracting with EtOAc. Combined organics washedwith brine, dried with sodium sulfate, filtered and concentrated invacuo. Crude residue was purified via silica gel column chromatographyto give Intermediate 71C (2.27 g, 8.86 mmol, 69% yield). TLC: productstains as purple spot in anisaldehyde (Rf=0.80 in 1:1 hex/EtOAc). ¹H NMR(400 MHz, chloroform-d) δ: 4.14 (q, J=7.5 Hz, 2H), 3.88-3.95 (m, 4H),2.09 (td, J=8.4, 5.6 Hz, 1H), 1.69-1.83 (m, 4H), 1.45-1.64 (m, 6H),1.33-1.42 (m, 1H), 1.25 (t, J=7.1 Hz, 3H), 0.86 (t, J=7.5 Hz, 3H).

Intermediate 71D. Ethyl 2-(4-oxocyclohexyl)butanoate

Intermediate 71C (2.00 g, 7.80 mmol) was taken up in THF (39.0 ml) andhydrochloric acid, 1M (39.0 ml) was added. Reaction stirred at roomtemperature for 2 hours. The reaction was concentrated in vacuo, dilutedwith water and extracted with EtOAc. The combined organic extracts weredried with sodium sulfate, filtered and concentrated in vacuo. The crudematerial was purified on silica gel column chromatography to giveIntermediate 71D (1.47 g, 6.92 mmol, 89% yield). TLC: product stainsfaintly pink in anisaldehyde (Rf=0.65 in 1:1 Hex/EtOAc). ¹H NMR (400MHz, chloroform-d) δ: 4.15 (q, J=7.1 Hz, 2H), 2.25-2.42 (m, 4H), 2.18(ddd, J=9.3, 7.8, 5.2 Hz, 1H), 2.10 (ddt, J=13.1, 6.2, 3.3 Hz, 1H),1.90-2.03 (m, 2H), 1.56-1.70 (m, 2H), 1.38-1.56 (m, 2H), 1.25 (t, J=7.2Hz, 3H), 0.89 (t, J=7.4 Hz, 3H).

Intermediate 71E. Ethyl 2-((trans)-4-hydroxycyclohexyl)butanoate

Intermediate 71D (1.47 g, 6.92 mmol) was dissolved in EtOH (13.85 ml)and cooled to 0° C. NaBH₄ (0.314 g, 8.31 mmol) was added and thereaction was then allowed to stir at 0° C. for 1 hour. The reaction wasquenched with saturated aqueous NH₄Cl and extracted with EtOAc. Combinedorganic extracts were dried with sodium sulfate, filtered, andconcentrated in vacuo. The crude material was purified via silica gelcolumn chromatography to give Intermediate 71E (1.22 g, 5.69 mmol, 82%yield) along with (138 mg, 0.644 mmol, 9.30% yield) of the cis-isomer.¹H NMR (400 MHz, chloroform-d) δ: 4.14 (q, J=7.1 Hz, 2H), 3.53 (t,J=10.5 Hz, 1H), 1.92-2.08 (m, 2H), 1.80-1.89 (m, 1H), 1.63-1.70 (m, 1H),1.52-1.62 (m, 4H), 1.37-1.52 (m, 2H), 1.26 (t, J=7.2 Hz, 3H), 0.95-1.17(m, 2H), 0.87 (t, J=7.4 Hz, 3H).

Intermediate 71F. Ethyl2-((trans)-4-(quinolin-4-yloxy)cyclohexyl)butanoate

Intermediate 71E (100 mg, 0.467 mmol) was taken up in DMSO (933 μl) andNaH (22.40 mg, 0.933 mmol) was added slowly, portionwise at rt. After 1hour, 4-bromoquinoline (117 mg, 0.560 mmol) was added and the reactionwas heated to 80° C. After 16 hours, the reaction was quenched withammonium chloride and extracted with EtOAc. The combined organicextracts were dried with sodium sulfate, filtered, concentrated invacuo. The crude residue was purified via silica gel columnchromatography to give Intermediate 71F (89 mg, 0.261 mmol, 55.9%yield). LC-MS Anal. Calc'd for C₂₁H₂₇NO₃ 341.20. found [M+H] 342.3T_(r)=0.84 min (Method A).

Intermediate 71G. 2-((trans)-4-(Quinolin-4-yloxy)cyclohexyl)butanoicacid

Intermediate 71F (89 mg, 0.261 mmol) was taken up in THF (1043 μl),Water (1043 μl), and MeOH (521 μl). Lithium hydroxide (62.4 mg, 2.61mmol) was added and reaction stirred at 60° C. for 24 hours. Thereaction was concentrated in vacuo, diluted with water, acidified withacetic acid added (precipitate forms), and extracted with EtOAc. Thecombined organic extracts were dried with sodium sulfate, filtered andconcentrated in vacuo to give Intermediate 71G (74 mg, 0.236 mmol, 91%yield. LC-MS Anal. Calc'd for C₁₉H₂₃NO₃ 313.17. found [M+H] 314.2T_(r)=0.69 min (Method A).

Intermediate 71H.1-((trans)-4-(Quinolin-4-yloxy)cyclohexyl)propan-1-amine

Intermediate 71G (190 mg, 0.606 mmol) was taken up in toluene (2021 μl)in a vial and diphenyl phosphorazidate (184 mg, 0.667 mmol) and TEA (101μl, 0.728 mmol) were added. The vial sealed and heated to 80° C. After 2h, the reaction was cooled to room temperature and concentrated underreduced pressure. The crude residue was taken up in 1 mL THF and 1 mL ofwater and LiOH (145 mg, 6.06 mmol) was added. Reaction stirred at roomtemperature for 1 hour. The reaction was acidified with 1N HCl (whiteprecipitate forms) and extracted with EtOAc to remove DPPA relatedimpurities. Then, the reaction was basified with 1N NaOH (precipitateforms again) and extracted with EtOAc (×5). Basic extracts wereconcentrated in vacuo to give Intermediate 71H (35 mg, 0.123 mmol,20.30% yield). LC-MS Anal. Calc'd for C₁₈H₂₄N₂O 284.19. found [M+H]285.2 T_(r)=0.55 min (Method A).

Example 714-Chloro-N-(1-((trans)-4-(quinolin-4-yloxy)cyclohexyl)propyl)benzamide

Intermediate 71H (35 mg, 0.123 mmol) was taken up in DMF (1231 μl) andHOBT (24.50 mg, 0.160 mmol), EDC (30.7 mg, 0.160 mmol), 4-chlorobenzoicacid (38.5 mg, 0.246 mmol) and TEA (86 μl, 0.615 mmol) were added andreaction was stirred at rt. After 2 hours, the reaction was diluted withDMF filtered and purified via preparative HPLC to give Example 71 (24.6mg, 0.058, 46.8% yield). LC-MS Anal. Calc'd for C₂₅H₂₇ClN₂O₂ 422.18.found [M+H] 423.3 T_(r)=0.82 min (Method A). ¹H NMR (500 MHz, DMSO-d₆)δ: 8.67 (d, J=5.2 Hz, 1H), 8.15 (d, J=9.0 Hz, 1H), 8.11 (d, J=8.2 Hz,1H), 7.87-7.94 (m, 3H), 7.72 (t, J=7.6 Hz, 1H), 7.49-7.58 (m, 3H), 7.10(d, J=5.2 Hz, 1H), 4.62 (t, J=10.2 Hz, 1H), 3.74-3.85 (m, J=8.9 Hz, 1H),2.21 (d, J=10.1 Hz, 2H), 1.86 (t, J=14.3 Hz, 2H), 1.39-1.71 (m, 5H),1.28 (q, J=12.3 Hz, 2H), 0.85 (t, J=7.2 Hz, 3H).

Enantiomer 1 and Enantiomer 2 Enantiomer 1 Example 714-Chloro-N-(1-((trans)-4-(quinolin-4-yloxy)cyclohexyl)propyl)benzamide(Homochiral, Stereochemistry Unknown)

Enantiomer 2 Example 714-Chloro-N-(1-((trans)-4-(quinolin-4-yloxy)cyclohexyl)propyl)benzamide(Homochiral, Stereochemistry Unknown)

Example 71 Enantiomer 1 and Enantiomer 2

Chiral separation of the racemic sample (Method C) gave Enantiomer 1T_(r)=5.195 min (Method D) and Enantiomer 2 T_(r)=8.226 min (Method D)Absolute stereochemistry was not determined.

Enantiomer 1: MS(ES): m/z=423.3 [M+H]⁺. T_(r)=2.126 min (Method A). ¹HNMR (500 MHz, DMSO-d₆) δ: 8.78 (d, J=5.1 Hz, 1H), 8.14-8.22 (m, 2H),7.97 (d, J=8.2 Hz, 1H), 7.89 (d, J=8.2 Hz, 2H), 7.82 (t, J=7.6 Hz, 1H),7.62 (t, J=7.5 Hz, 1H), 7.54 (d, J=8.3 Hz, 2H), 7.25 (d, J=5.6 Hz, 1H),4.65-4.75 (m, 1H), 3.75-3.84 (m, J=8.8 Hz, 1H), 2.22 (d, J=10.4 Hz, 2H),1.81-1.92 (m, 2H), 1.43-1.70 (m, 5H), 1.28 (q, J=12.2 Hz, 2H), 0.85 (t,J=7.2 Hz, 3H).

Enantiomer 2: MS(ES): m/z=423.3[M+H]⁺. T_(r)=2.126 min (Method A). ¹HNMR (500 MHz, DMSO-d₆) δ: 8.67 (d, J=5.0 Hz, 1H), 8.16 (d, J=9.0 Hz,1H), 8.11 (d, J=8.2 Hz, 1H), 7.85-7.94 (m, 3H), 7.72 (t, J=7.3 Hz, 1H),7.54 (d, J=8.2 Hz, 3H), 7.09 (d, J=5.2 Hz, 1H), 4.61 (t, J=10.1 Hz, 1H),3.75-3.83 (m, J=8.5 Hz, 1H), 2.21 (d, J=10.2 Hz, 2H), 1.85 (t, J=14.0Hz, 2H), 1.39-1.69 (m, 5H), 1.22-1.33 (m, 2H), 0.85 (t, J=7.1 Hz, 3H).

Example 724-Chloro-N-(1-((trans)-4-((8-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)propyl)benzamide

Example 72 was prepared from Intermediate 71E and the analogousprocedures outlined to make 71F, 71G, 71H, and Example 71 except that4-chloro-8-(trifluoromethyl)quinoline was used in part F. LC-MS Anal.Calc'd for C₂₆H₂₆ClF₃N₂O₂ 490.16. found [M+H] 491.2 T_(r)=0.99 min(Method A). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.80 (d, J=5.0 Hz, 1H), 8.40(d, J=8.2 Hz, 1H), 8.17 (d, J=8.9 Hz, 1H), 8.14 (d, J=7.2 Hz, 1H), 7.88(d, J=8.2 Hz, 2H), 7.65 (t, J=7.8 Hz, 1H), 7.53 (d, J=8.2 Hz, 2H), 7.24(d, J=5.1 Hz, 1H), 4.66 (t, J=10.0 Hz, 1H), 3.74-3.84 (m, J=6.6 Hz, 1H),2.21 (d, J=10.2 Hz, 2H), 1.85 (t, J=13.5 Hz, 2H), 1.40-1.71 (m, 5H),1.27 (q, J=12.1 Hz, 2H), 0.84 (t, J=7.0 Hz, 3H).

Example 73 and Example 74(trans)-N-(4-Chlorobenzyl)-4-(6-fluoroquinolin-4-yl)cyclohexanecarboxamide(cis)-N-(4-Chlorobenzyl)-4-(6-fluoroquinolin-4-yl)cyclohexanecarboxamide(Homochiral, Absolute and Relative Stereochemistry Unassigned)

Intermediate 73A. 1,4-Dioxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate

To a stirred solution of 1,4-dioxaspiro[4.5]decan-8-one (300 g, 1920.86mmol, 1.0 eq) and N-phenyltrifluoromethanesulfonimide (823.47 g, 2305.03mmol, 1.2 eq) in MTBE (7.5 L) under nitrogen atmosphere at −78° C. wasadded 2.0 M NaHMDS in THF (1152.2 mL, 2305.03 mmol, 1.2 eq) over 70minutes, and the mixture was stirred for an additional 60 minutes. Thereaction mixture was warmed to room temperature and stirred overnightuntil TLC showed complete consumption of the starting material. Themixture was quenched with aqueous KHSO₄ (100 ml), filtrated to removethe solid and concentrated the filtrate completely. To the residue wasadded 3 L MTBE, then washed with 5% NaOH (1.5 L×3). The organic phasewas concentrated to obtain Intermediate 34A (567 g, light yellow oil,yield 102% yield). TLC Rf: 0.7 (PE/EtOAc=10/1, KMnO₄). ¹H NMR (400 MHz,CDCl₃): δ (ppm) 5.65 (t, J=4.0 Hz, 1H), 3.98 (d, J=1.5 Hz, 4H), 2.53 (s,2H), 2.40 (s, 2H), 1.90 (t, J=6.6 Hz, 2H).

Intermediate 73B.4,4,5,5-Tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane

A mixture of Intermediate 73A (600 g, 2.08 mol, 1 eq), B₂Pin₂ (687.1 g,2.71 mol, 1.3 eq), KOAc (613 g, 6.24 mol, 3 eq), NaBr (86 g 0.833 mol,0.4 eq) and Pd(dppf)Cl₂ (76 g, 0.1 mol, 0.05 eq) in dioxane (6.5 L) washeated to reflux overnight. Once the reaction was complete, the mixturewas concentrated and purified by via silica gel column chromatography togive Intermediate 73B (369 g, 66% yield). LC-MS Anal. Calc'd forC₁₄H₂₃BO₄ 266.17. found [M+H] 267.1. ¹H NMR (400 MHz, CDCl₃) δ 6.46 (s,1H), 3.98 (s, 4H), 2.37-2.35 (m, 4H), 1.74-1.60 (t, 2H), 1.24 (s, 12H).

Intermediate 73C. 6-Fluoro-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)quinoline

A mixture of Intermediate 73B (368 g, 1.38 mol, 1.3 eq),4-chloro-6-fluoroquinoline (195 g, 1.07 mol, 1 eq), K₂CO₃ (445 g, 3.22mol, 3 eq) and Pd(PPh₃)₄ (25 g, 22 mmol, 0.02 eq) in dioxane-water (3 L,4:1) was heated to reflux overnight. The solution was concentrated andextracted with EtOAc. The crude residue was purified via silica gelcolumn chromatography to give Intermediate 73C (236 g, 77% yield). LC-MSAnal. Calc'd for C₁₇H₁₆FNO₂ 285.12. found [M+H] 286.1. ¹H NMR (400 MHz,CDCl₃) δ 8.80-8.29 (d, 1H), 8.11-8.07 (q, 1H), 7.63-7.61 (q, 1H),7.47-7.46 (q, 1H), 7.26-7.22 (m, 1H), 5.75-5.74 (m, 1H), 4.08-4.05 (m,4H), 2.63-2.59 (m, 2H), 2.59-2.53 (m, 2H), 2.0-1.97 (m, 2H).

Intermediate 73D. 6-Fluoro-4-(1,4-dioxaspiro[4.5]decan-8-yl)quinoline

To Intermediate 73C (125 g, 0.44 mol) in IPA (2 L) at 55° C. was added10% Pd/C and the mixture was stirred under an atmosphere of H₂overnight. The mixture was filtered and concentrated to give crudeIntermediate 73D (128 g, 100% yield), which was used directly in thenext step. LC-MS Anal. Calc'd for C₁₇H₁₈FNO₂ 287.13. found [M+H] 288.2,rt=0.62 min (Method A).

Intermediate 73E. 4-(6-Fluoroquinolin-4-yl)cyclohexanone

Intermediate 73D (100 g, 0.348 mol) was treated with 4 N HCl (300 mL) inacetone (1200 mL) at 45° C. for overnight. Then the solution wasconcentrated in vacuo. The residue was adjusted pH 9 with 6 N NaOH. Themixture was partitioned between ethyl acetate and water. The organiclayer was washed with brine, dried over anhydrous Na₂SO₄, filtered andconcentrated to give light yellow solid, which was then purified bysilica gel column chromatography to afford Intermediate 73E as whitesolid (67 g, 55% yield). LC-MS Anal. Calc'd for C₁₅H₄FNO 243.11. found[M+H] 244.0. ¹H NMR (400 MHz, CDCl₃) δ 8.84 (d, J=4.6 Hz, 1H), 8.16 (dd,J=9.3, 5.7 Hz, 1H), 7.72 (dd, J=10.3, 2.8 Hz, 1H), 7.52 (ddd, J=9.2,7.8, 2.7 Hz, 1H), 7.29 (d, J=4.6 Hz, 1H), 3.69 (ddd, J=12.1, 9.0, 3.3Hz, 1H), 2.77-2.54 (m, 4H), 2.37 (ddd, J=13.4, 5.9, 3.0 Hz, 2H), 2.04(qd, J=12.6, 5.3 Hz, 2H).

Intermediate 73F. 4-(6-Fluoroquinolin-4-yl)cyclohexanecarbonitrile

To a solution of Intermediate 73E (500 mg, 2.055 mmol) and1-((isocyanomethyl)sulfonyl)-4-methylbenzene (522 mg, 2.67 mmol) in DMSO(10.100 ml) and methanol (0.202 ml) was added potassium2-methylpropan-2-olate (554 mg, 4.93 mmol). After the addition wascomplete, the reaction mixture was stirred room temperature. After 1hour, the reaction was diluted with diethyl ether (30 mL) and washedwith water (20 ml). The organic layer was dried with anhydrous MgSO₄,concentrated under reduced pressure. The crude material was purified viasilica gel column chromatography to give Intermediate 73F (280 mg, 1.101mmol, 53.6% yield) as a mixture of cis and trans isomers (˜2:1 ratio).LC-MS Anal. Calc'd for C₁₆H₁₅FN₂ 254.12. found [M+H] 255.1, rt=0.60(first elution diastereomer) and 0.62 min (second eluting diastereomer)(Method A).

Intermediate 73G. 4-(6-Fluoroquinolin-4-yl)cyclohexanecarboxylic acid

Intermediate 73F (280 mg, 1.101 mmol) taken up in methanol (5505 μl) andhydrochloric acid, 37% (5505 μl). Reaction heated at 70° C. After 48hours, the reaction was slowly added to 100 mL water and basified withsodium bicarbonate (sat aq). The aqueous was extracted with EtOAc. Thecombined organics were dried with sodium sulfate, filtered, andconcentrated in vacuo to give crude methyl4-(6-fluoroquinolin-4-yl)cyclohexanecarboxylate. This crude material wastaken up in THF (4260 μl), Water (4260 μl), MeOH (2130 μl) and lithiumhydroxide (255 mg, 10.65 mmol) was added. Reaction stirred at roomtemperature for 1 hour. The reaction was then concentrated, acidifiedwith 1N HCl and extracted with EtOAc. The combined organics were driedwith sodium sulfate, filtered, and concentrated in vacuo to give a cruderesidue. This crude residue was purified via silica gel columnchromatography to give Intermediate 73G (mixture of cis and trans) (63mg, 0.231 mmol, 21.64% yield). LC-MS Anal. Calc'd for C₁₆H₁₆FNO₂ 273.1.found [M+H] 274.1, rt=0.55 (Method A).

Example 73 and Example 74N-(4-Chlorobenzyl)-4-(6-fluoroquinolin-4-yl)cyclohexanecarboxamide(Mixture of Cis- and Trans-Isomers)

Intermediate 73G (15 mg, 0.055 mmol) was dissolved in thionyl chloride(40.1 μl, 0.549 mmol) and DMF (2.125 μl, 0.027 mmol) was added. Reactionstirred at room temperature for 1 hour. After, the reaction wasconcentrated in vacuo, taken up in toluene, concentrated again andplaced on high vac. After 15 minutes, the crude acyl chloride was takenup in ACN (274 μl) and added to a solution of(4-chlorophenyl)methanamine (15.54 mg, 0.110 mmol) in ACN (274 μl) andTEA (38.2 μl, 0.274 mmol) at 0° C. Reaction was then allowed to warm toroom temperature. After 1 hour, the reaction was diluted with water andextracted with EtOAc. Organics were dried with sodium sulfate, filtered,and concentrated in vacuo. Crude residue taking up in DMF filtered, andpurified via preparative HPLC to give two diastereomers.

The first eluting diastereomer, Example 73 (4.9 mg, 0.012 mmol, 22%yield). LC-MS Anal. Calc'd for C₂₃H₂₂ClFN₂O 396.14. found [M+H] 397.0,rt=1.891 (Method B). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.80 (d, J=4.4 Hz,1H), 8.40 (t, J=5.7 Hz, 1H), 8.08 (dd, J=9.0, 5.9 Hz, 1H), 7.99 (d,J=9.8 Hz, 1H), 7.66 (t, J=8.6 Hz, 1H), 7.45 (d, J=4.3 Hz, 1H), 7.37 (d,J=8.2 Hz, 2H), 7.26 (d, J=8.1 Hz, 2H), 4.26 (d, J=5.8 Hz, 2H), 3.33 (t,J=11.9 Hz, 1H), 2.32 (t, J=11.9 Hz, 1H), 1.92 (t, J=11.2 Hz, 4H),1.71-1.83 (m, 2H), 1.56 (q, J=12.3 Hz, 2H).

The second eluting enantiomer, Example 74 (3.6 mg, 0.009 mmol, 17%yield) LC-MS Anal. Calc'd for C₂₃H₂₂ClFN₂O 396.14. found [M+H] 397.0,rt=1.940 (Method B). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.78 (d, J=4.3 Hz,1H), 8.37 (t, J=5.5 Hz, 1H), 8.07 (dd, J=8.9, 6.0 Hz, 1H), 7.95 (d,J=10.8 Hz, 1H), 7.65 (t, J=8.6 Hz, 1H), 7.36 (d, J=8.0 Hz, 2H),7.23-7.31 (m, 3H), 4.27 (d, J=5.7 Hz, 2H), 3.33 (br. s., 1H), 2.61 (br.s., 1H), 2.01-2.13 (m, 2H), 1.74-1.85 (m, 4H), 1.65-1.74 (m, 2H).

Example 75 and Example 76(trans)-N-(4-Chlorophenyl)-4-(6-fluoroquinolin-4-yl)cyclohexanecarboxamide(cis)-N-(4-Chlorophenyl)-4-(6-fluoroquinolin-4-yl)cyclohexanecarboxamide(Homochiral, Absolute and Relative Stereochemistry Unassigned)

Example 75 and Example 76 were made from Intermediate 73G utilizing theprocedure to make Example 73 and Example 74.

First eluting diastereomer: (7.1 mg, 0.018 mmol, 34% yield) LC-MS Anal.Calc'd for C₂₂H₂₀ClFN₂O 382.13. found [M+H] 383.2, rt=2.011 (Method B).¹H NMR (500 MHz, DMSO-d₆) δ: 10.00 (s, 1H), 8.78 (d, J=4.4 Hz, 1H), 8.07(dd, J=9.1, 5.8 Hz, 1H), 7.96 (d, J=8.8 Hz, 1H), 7.59-7.68 (m, 3H), 7.39(d, J=4.4 Hz, 1H), 7.33 (d, J=8.8 Hz, 2H), 3.37 (br. s., 1H), 2.78 (br.s., 1H), 2.09 (d, J=12.1 Hz, 2H), 1.81-2.00 (m, 4H), 1.75 (d, J=11.4 Hz,2H).

Second eluting diastereomer: (8.4 mg, 0.021 mmol, 39% yield) LC-MS Anal.Calc'd for C₂₂H₂₀ClFN₂O 382.13. found [M+H] 383.0, rt=1.988 (Method B).¹H NMR (500 MHz, DMSO-d₆) δ: 10.09 (s, 1H), 8.81 (d, J=4.4 Hz, 1H), 8.08(dd, J=9.0, 5.9 Hz, 1H), 8.01 (d, J=8.9 Hz, 1H), 7.61-7.71 (m, 3H), 7.46(d, J=4.4 Hz, 1H), 7.34 (d, J=8.7 Hz, 2H), 3.36 (t, J=11.9 Hz, 1H),2.41-2.48 (m, 1H) (triplet buried under DMSO), 1.97 (d, J=9.7 Hz, 4H),1.76-1.88 (m, 2H), 1.54-1.65 (m, 2H).

Example 77a(±)-4-Chloro-N-(1-((cis)-4-(pyridin-4-yloxy)cyclohexyl)propyl)benzamide

Intermediate 77A. Ethyl 2-((cis)-4-(pyridin-4-yloxy)cyclohexyl)butanoate

Intermediate 71E (100 mg, 0.467 mmol) was dissolved in THF (1867 μl) andpyridin-4-ol (98 mg, 1.027 mmol) and triphenylphosphine (269 mg, 1.027mmol) were added. Solution was cooled to 0° C. in an ice bath.Diisopropyl azodicarboxylate (200 μl, 1.027 mmol) was added and thereaction was allowed to stir at room temperature once the addition wascomplete. Stirred at room temperature for 16 hours. Reaction wasconcentrated in vacuo and purified via silica gel column chromatographyto give Intermediate 77A (89 mg, 0.205 mmol, 43.9% yield). LC-MS Anal.Calc'd for C₁₇H₂₅NO₃ 291.18. found [M+H] 292.3 T_(r)=0.84 min (MethodA). ¹H NMR (400 MHz, chloroform-d) δ: 8.34-8.42 (m, 2H), 6.71-6.79 (m,2H), 4.57-4.64 (m, 1H), 4.15 (q, J=7.1 Hz, 2H), 2.14 (ddd, J=9.8, 7.9,4.6 Hz, 1H), 1.97-2.07 (m, 2H), 1.38-1.69 (m, 9H), 1.24-1.29 (m, 3H),0.88 (t, J=7.4 Hz, 3H)

Intermediate 77B. 2-((cis)-4-(Pyridin-4-yloxy)cyclohexyl)butanoic acid

Intermediate 77A (89 mg, 0.305 mmol) was taken up in THF (244 μl), water(244 μl), and MeOH (122 μl). Lithium hydroxide (73.1 mg, 3.05 mmol) wasadded and the reaction stirred at 60° C. for 16 hours. Lithium hydroxide(73.1 mg, 3.05 mmol) was again added and the reaction stirred foranother 24 hours at 60° C. The reaction was concentrated in vacuo,diluted with water and extracted with EtOAc. The aqueous layer was thentreated with AcOH and extracted with EtOAc. LCMS shows Product remainsin aqueous layer. Extracted again with 7:3 chloroform:propanol. LCMSShows product was successfully extracted from the aqueous layer. Thecombined organic layers were dried with sodium sulfate, filtered, andconcentrated in vacuo to give Intermediate 77B (73 mg, 0.277 mmol, 91%yield). Material was not further purified. LC-MS Anal. Calc'd forC₁₅H₂₁NO₃ 263.15. found [M+H] 264.2 T_(r)=0.58 min (Method A).

Intermediate 77C. 1-((cis)-4-(Pyridin-4-yloxy)cyclohexyl)propan-1-amine

Intermediate 77B (35 mg, 0.133 mmol) was taken up in toluene (443 μl)and diphenyl phosphorazidate (40.2 mg, 0.146 mmol) and TEA (22.23 μl,0.159 mmol) added. The reaction vial was sealed (vented with needlewhile reaching temperature) and heated to 80° C. After 2 h, the reactionwas cooled to room temperature and concentrated under reduced pressure.The crude residue was taken up in 1 mL THF and 1 mL of water LiOH (31.8mg, 1.329 mmol) was added. The reaction stirred at room temperatureovernight. The reaction was acidified with 1N HCl and extracted withEtOAc (extracts discarded). The aqueous portion was then basified with1N NaOH and extracted with EtOAc (×2). The combined basic, organicextracts were dried with sodium sulfate, filtered and concentrated invacuo to give Intermediate 77C (25 mg, 0.107 mmol, 80% yield). LC-MSAnal. Calc'd for C₁₄H₂₂N₂O 234.17. found [M+H] 235.1 T_(r)=0.43 min(Method A).

Example 77a(±)-4-Chloro-N-(1-((cis)-4-(pyridin-4-yloxy)cyclohexyl)propyl)benzamide

Intermediate 77C (25 mg, 0.107 mmol) was taken up in DMF (1067 μl) andHOBT (21.24 mg, 0.139 mmol), EDC (26.6 mg, 0.139 mmol), 4-chlorobenzoicacid (33.4 mg, 0.213 mmol) and TEA (74.3 μl, 0.533 mmol) were added andreaction stirred at room temperature. After 2 hours, the reaction wasdiluted with DMF to bring total volume to 2 mL, filtered, and purifiedvia preparative HPLC to give Example 77a (23.2 mg, 0.062 mmol, 58%yield). LC-MS Anal. Calc'd for C₂₁H₂₅ClN₂O₂ 372.16. found [M+H] 373.3T_(r)=0.73 min (Method A). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.30 (d, J=5.6Hz, 2H), 8.17 (d, J=9.1 Hz, 1H), 7.81 (d, J=8.3 Hz, 2H), 7.51 (d, J=8.3Hz, 2H), 6.91 (d, J=5.6 Hz, 2H), 4.69 (br. s., 1H), 1.88 (d, J=12.0 Hz,2H), 1.25-1.68 (m, 9H), 0.81 (t, J=7.2 Hz, 3H).

Examples 77b and c4-Chloro-N-((R)-1-((cis)-4-(pyridin-4-yloxy)cyclohexyl)propyl)benzamide4-Chloro-N-((S)-1-((cis)-4-(pyridin-4-yloxy)cyclohexyl)propyl)benzamide(Homochiral, Absolute and Relative Stereochemistry Unassigned)

Example 77b and Example 77c

Chiral separation of the racemic Example 77a (Method E) gave Example 77bT_(r)=3.391 min (Method F) and Example 77c T_(r)=3.851 min (Method F)Absolute stereochemistry was not determined.

Example 77b

MS(ES): m/z=373.3 [M+H]⁺. T_(r)=1.783 min (Method B). ¹H NMR (500 MHz,DMSO-d₆) δ: 8.34 (d, J=4.9 Hz, 2H), 8.12 (d, J=9.0 Hz, 1H), 7.84 (d,J=8.3 Hz, 2H), 7.53 (d, J=8.3 Hz, 2H), 6.94 (d, J=5.5 Hz, 2H), 4.71 (br.s., 1H), 1.89 (br. s., 2H), 1.27-1.69 (m, 10H), 0.83 (t, J=7.2 Hz, 3H).

Example 77c

MS(ES): m/z=373.3 [M+H]⁺. T_(r)=1.793 min (Method B). ¹H NMR (500 MHz,DMSO-d₆) δ: 8.34 (d, J=3.6 Hz, 2H), 8.12 (d, J=9.0 Hz, 1H), 7.84 (d,J=8.3 Hz, 2H), 7.53 (d, J=8.4 Hz, 2H), 6.95 (d, J=5.4 Hz, 2H), 4.72 (br.s., 1H), 1.86-1.94 (m, J=5.0 Hz, 2H), 1.29-1.68 (m, 10H), 0.83 (t, J=7.2Hz, 3H).

Example 78(±)-4-Chloro-N-(1-((cis)-4-((6-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)propyl)benzamide

Example 78 was synthesized from Intermediate 71E following the sameprocedures used to make Intermediates 77A, 77B, 77C and Example 77ausing 4-hydroxy-6-trifluoromethyl quinoline rather than pyridin-4-ol inthe synthesis of 77A. LC-MS Anal. Calc'd for C₂₆H₂₆ClF₃N₂O₂ 490.16.found [M+H] 491.2 T_(r)=0.86 min (Method A). ¹H NMR (500 MHz, DMSO-d₆)δ: 8.81 (d, J=5.2 Hz, 1H), 8.36 (s, 1H), 8.16 (d, J=9.0 Hz, 1H), 8.12(d, J=8.8 Hz, 1H), 7.96 (d, J=8.2 Hz, 1H), 7.81 (d, J=8.3 Hz, 2H), 7.45(d, J=8.4 Hz, 2H), 7.15 (d, J=5.3 Hz, 1H), 4.98 (br. s., 1H), 2.07 (t,J=15.6 Hz, 2H), 1.37-1.74 (m, 9H), 0.82 (t, J=7.2 Hz, 3H).

Example 79a(±)-4-Chloro-N-(1-((cis)-4-((2-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)propyl)benzamide

Example 79a was synthesized from Intermediate 71E following the sameprocedures used to make Intermediates 77A, 77B, 77C and Example 77ausing 4-hydroxy-2-trifluoromethyl quinoline rather than pyridin-4-ol inthe synthesis of 77A. LC-MS Anal. Calc'd for C₂₆H₂₆ClF₃N₂O₂ 490.16.found [M+H] 491.2 T_(r)=1.13 min (Method A). ¹H NMR (500 MHz, DMSO-d₆)δ: 8.17 (d, J=8.3 Hz, 1H), 8.14 (d, J=9.1 Hz, 1H), 8.05 (d, J=8.4 Hz,1H), 7.81-7.91 (m, 3H), 7.50-7.58 (m, 3H), 7.39 (s, 1H), 5.18 (br. s.,1H), 2.07 (t, J=13.9 Hz, 2H), 1.40-1.74 (m, 9H), 0.85 (t, J=7.2 Hz, 3H).

Examples 79b and c4-Chloro-N-((R)-1-((cis)-4-(2-(trifluoromethyl)quinolin-4-yloxy)cyclohexyl)propyl)benzamideand4-Chloro-N-((S)-1-((cis)-4-(2-(trifluoromethyl)quinolin-4-yloxy)cyclohexyl)propyl)benzamide(Homochiral, Absolute and Relative Stereochemistry Unassigned)

Example 79b and Example 79c

Chiral separation of the racemic Example 79a (Method G) gave Example 79bT_(r)=3.998 min (Method H) and Example 79c T_(r)=5.009 min (Method H)Absolute stereochemistry was not determined.

Example 79b

MS(ES): m/z=491.3 [M+H]⁺. T_(r)=2.438 min (Method B). ¹H NMR (500 MHz,DMSO-d₆) δ: 8.11-8.20 (m, 2H), 0.8.05 (d, J=8.3 Hz, 1H), 7.81-7.89 (m,J=7.0 Hz, 3H), 7.49-7.57 (m, 3H), 7.37 (s, 1H), 5.16 (br. s., 1H), 3.83(br. s., 1H), 2.06 (t, J=13.4 Hz, 2H), 1.56-1.76 (m, 6H), 1.39-1.56 (m,3H), 0.84 (t, J=6.9 Hz, 3H)

Example 79c

MS(ES): m/z=491.3 [M+H]⁺. T_(r)=2.438 min (Method B). ¹H NMR (500 MHz,DMSO-d₆) δ: 8.16 (t, J=8.8 Hz, 2H), 8.04 (d, J=8.5 Hz, 1H), 7.85 (m,3H), 7.49-7.56 (m, 3H), 7.37 (s, 1H), 5.15 (br. s., 1H), 3.83 (br. s.,1H), 2.00-2.11 (m, 2H), 1.56-1.73 (m, 6H), 1.40-1.55 (m, 3H), 0.84 (t,J=7.2 Hz, 3H)

Example 804-Chloro-N-(1-(cis-4-(quinolin-4-yloxy)cyclohexyl)propyl)benzamide

Example 80 was synthesized from Intermediate 71E following the sameprocedures used to make Intermediates 77A, 77B, 77C and Example 77 using4-hydroxyquinoline rather than pyridin-4-ol as in the synthesis of 77A.LC-MS Anal. Calc'd for C₂₅H₂₇ClN₂O₂ 422.18. found [M+H] 423.2 T_(r)=0.78min (Method A). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.66 (d, J=5.0 Hz, 1H),8.14 (d, J=9.0 Hz, 1H), 8.08 (d, J=8.1 Hz, 1H), 7.90 (d, J=8.3 Hz, 1H),7.85 (d, J=8.2 Hz, 2H), 7.70 (t, J=7.4 Hz, 1H), 7.52 (d, J=8.2 Hz, 2H),7.36 (t, J=7.5 Hz, 1H), 7.00 (d, J=5.1 Hz, 1H), 4.95 (br. s., 1H), 2.06(t, J=13.9 Hz, 2H), 1.38-1.71 (m, 9H), 0.84 (t, J=7.0 Hz, 3H).

Example 814-Chloro-N-((1-(6-fluoroquinolin-4-yl)-4-hydroxypiperidin-4-yl)methyl)benzamide

81A. tert-Butyl4-((4-chlorobenzamido)methyl)-4-hydroxypiperidine-1-carboxylate

A solution of tert-butyl4-(aminomethyl)-4-hydroxypiperidine-1-carboxylate (0.25 g, 1.086 mmol)and 4-chlorobenzoic acid (0.204 g, 1.303 mmol) in DMF (2 mL) was treatedwith triethylamine (0.454 mL, 3.26 mmol) followed by BOP (0.576 g, 1.303mmol). The reaction was stirred for 2 h then quenched with dil. aq.HOAc. This resulted in the formation of a precipitate, so the mixturewas filtered and rinsed with water. It was then suspended in dil. aq.sodium bicarbonate, sonicated, then filtered, rinsed with water, andair-dried to afford tert-butyl4-((4-chlorobenzamido)methyl)-4-hydroxypiperidine-1-carboxylate (0.38 g,90% yield) as a colorless solid, mp 172-173° C. MS(ES): m/z=369 [M+H]⁺.t_(R)=0.93 min (Method A).

81B. 4-Chloro-N-((4-hydroxypiperidin-4-yl)methyl)benzamide, HCl

A solution of HCl (3.56 ml, 14.23 mmol) in dioxane was treated withtert-butyl4-((4-chlorobenzamido)methyl)-4-hydroxypiperidine-1-carboxylate (0.35 g,0.949 mmol). Initially, material dissolved as it was added, buteventually, a gum formed. This was stirred for 15 min. then ˜3 mL ofdichloromethane was added and stirring was continued for 2 h. Duringthis time, product took on the form of a finely-divided suspension.Concentration under reduced pressure afforded4-chloro-N-((4-hydroxypiperidin-4-yl)methyl)benzamide, HCl (0.29 g,quantitative yield) as a white powder. MS(ES): m/z=269 [M+H]⁺.t_(R)=0.55 min (Method A).

Example 814-Chloro-N-((1-(6-fluoroquinolin-4-yl)-4-hydroxypiperidin-4-yl)methyl)benzamide

A suspension of 4-chloro-6-fluoroquinoline (0.119 g, 0.655 mmol) and4-chloro-N-((4-hydroxypiperidin-4-yl)methyl)benzamide, HCl (0.2 g, 0.655mmol) in NMP (1 mL) was treated with DIEA (0.286 mL, 1.638 mmol) andheated to 135° C. for 5 h. After about 45 min. the reaction had becomehomogeneous. The reaction was cooled to ˜80° C. and treated with ˜3 mLof 5% aq. HOAc resulting in the formation of a precipitate. This wasstirred briefly, filtered, rinsed several times with water and once with10% EtOAc-hexanes, and air-dried to afford4-chloro-N-((1-(6-fluoroquinolin-4-yl)-4-hydroxypiperidin-4-yl)methyl)benzamide(0.21 g, 74% yield) as an off-white solid, mp 91-94° C. MS(ES): m/z=414[M+H]⁺. t_(R)=0.68 min (Method A). ¹H NMR (400 MHz, DMSO-d₆) δ 8.67 (d,1H, J=4.9 Hz), 8.46 (t, 1H, J=6.1 Hz), 7.99-8.04 (m, 1H), 7.94 (d, 2H,J=8.7 Hz), 7.55-7.63 (m, 4H), 7.05 (d, 1H, J=5.0 Hz), 4.71 (s, 1H), 3.42(d, 2H, J=6.1 Hz), 3.24-3.31 (m, integration obscured by water peak),3.10-3.18 (m, 2H), 1.85-1.94 (m, 2H), 1.67-1.72 (m, 2H).

Example 84N-([1,1′-Biphenyl]-4-yl)-4-(6-fluoroquinolin-4-yl)piperazine-1-carboxamide

84A. tert-Butyl 4-(6-fluoroquinolin-4-yl)piperazine-1-carboxylate

To a homogeneous mixture of 4-chloro-6-fluoroquinoline (500.0 mg, 2.8mmol) in NMP (5 mL), in a sealable vial, was added 1-Boc-piperazine(750.0 mg, 4.0 mmol) followed by DIPEA (2 mL, 11.6 mmol). After theaddition was complete, the vial was capped and the mixture was stirredat 120° C. After 15 hours, the reaction was cooled to room temperaturethen partitioned between water and Et₂O. The layers were separated andthe aqueous layer was extracted twice more with Et₂O then once withEtOAc. These organic extracts were combined with the original organiclayer and were washed with brine, dried (Na₂SO₄), filtered andconcentrated in vacuo to afford the crude product as an oil.Purification by Isco chromatography afforded tert-butyl4-(6-fluoroquinolin-4-yl)piperazine-1-carboxylate as a solid (719.3 mg;77% yield). MS(ES): m/z=332 [M+H]⁺. t_(R)=0.70 min (Method A). ¹H NMR(400 MHz, DMSO-d₆) δ 8.70 (d, J=4.9 Hz, 1H), 8.04 (dd, J=9.2, 5.6 Hz,1H), 7.76-7.58 (m, 2H), 7.07 (d, J=5.0 Hz, 1H), 3.71-3.54 (m, 4H),3.14-3.01 (m, 4H), 1.44 (s, 9H).

84B. 6-Fluoro-4-(piperazin-1-yl)quinoline

To a homogeneous mixture of tert-butyl4-(6-fluoroquinolin-4-yl)piperazine-1-carboxylate (700.0 mg, 2.1 mmol)in anhydrous dioxane (4 mL), at room temperature under nitrogen, wasadded HCl (4N in dioxane, 10 mL, 40.0 mmol). After 6 hours, aprecipitate had formed which was isolated by vacuum filtration, rinsedwith anhydrous dioxane and dried under vacuum to afford the titlecompound (508.0 mg, 79% yield) as an HCl salt which was used withoutfurther purification. MS(ES): m/z=232 [M+H]⁺. t_(R)=0.38 min (Method A).

84N-([1,1′-Biphenyl]-4-yl)-4-(6-fluoroquinolin-4-yl)piperazine-1-carboxamide

To a heterogeneous mixture of the HCl salt of6-fluoro-4-(piperazin-1-yl)quinoline (84B, 25.0 mg, 0.09 mmol) inanhydrous DMF (1 mL), at room temperature, was added DIPEA (0.05 mL,0.29 mmol) followed by 4-isocyanato-1,1′-biphenyl (23.0 mg, 0.12 mmol).The resulting mixture was stirred for 96 hours, before being dilutedwith DMF, passed through a syringe filter, then purified via preparativeHPLC/MS to afford the title compound (12.9 mg; 21% yield). MS(ES):m/z=427 [M+H]⁺. t_(R)=1.55 min (Method I). ¹H NMR (500 MHz, DMSO-d₆) δ8.69 (d, J=6.5 Hz, 1H), 8.07 (dd, J=9.2, 5.1 Hz, 1H), 7.99-7.96 (m, 1H),7.93-7.90 (m, 1H), 7.63 (d, J=7.7 Hz, 2H), 7.58-7.57 (m, 4H), 7.44-7.41(m, 2H), 7.33-7.26 (m, 2H), 7.20 (d, J=6.6 Hz, 1H), 3.87-3.81 (m, 4H),3.81-3.73 (m, 4H).

Example 86N-((1-(6-Fluoroquinolin-4-yl)piperidin-4-yl)methyl)-4-methylbenzamide

86A. tert-Butyl((1-(6-fluoroquinolin-4-yl)piperidin-4-yl)methyl)carbamate

To a homogeneous mixture of 4-chloro-6-fluoroquinoline (220.0 mg, 1.2mmol) in anhydrous NMP (4 mL), in a sealable vial, was added tert-butyl(piperidin-4-ylmethyl)carbamate (350.0 mg, 1.6 mmol) followed by DIPEA(0.8 mL, 4.6 mmol). The vial was sealed and the mixture was stirred at60° C. for 2 hours, then at 90° C. for 17 hours before being stirred at120° C. for 24 hours. After cooling to room temperature, the reactionmixture was purified by Isco silica gel chromatography to affordtert-butyl ((1-(6-fluoroquinolin-4-yl)piperidin-4-yl)methyl)carbamate asan off-white solid (323.7 mg; 74% yield). MS(ES): m/z=360 [M+H]⁺.t_(R)=0.71 min (Method A). ¹H NMR (400 MHz, DMSO-d₆) δ 8.66 (d, J=5.0Hz, 1H), 8.01 (dd, J=9.1, 5.7 Hz, 1H), 7.66-7.51 (m, 2H), 7.01 (d, J=4.9Hz, 1H), 6.93 (t, J=5.7 Hz, 1H), 3.48 (d, J=12.2 Hz, 2H), 2.94 (t, J=6.3Hz, 2H), 2.76 (t, J=11.2 Hz, 2H), 1.80 (d, J=11.1 Hz, 2H), 1.67-1.55 (m,1H), 1.51-1.42 (m, 2H), 1.40 (s, 9H).

86B. (1-(6-Fluoroquinolin-4-yl)piperidin-4-yl)methanamine

To a homogeneous mixture of tert-butyl((1-(6-fluoroquinolin-4-yl)piperidin-4-yl)methyl)carbamate (308.1 mg,0.9 mmol) in DCM (10 mL), under nitrogen atmosphere, was added TFA (1.2mL, 15.6 mmol). The resultant mixture was stirred at ambient temperaturefor 45 minutes before being concentrated in vacuo to afford the TFA saltof the title compound as a gold oil, which was used without furtherpurification. MS(ES): m/z=260 [M+H]⁺. t_(R)=0.43 min (Method A).

Example 86N-((1-(6-Fluoroquinolin-4-yl)piperidin-4-yl)methyl)-4-methylbenzamide

To a homogeneous mixture of the TFA salt of(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)methanamine (86B, 41.8 mg, 0.09mmol), 4-methylbenzoic acid (14.0 mg, 0.1 mmol) and DIPEA (0.06 mL, 0.3mmol) in anhydrous THF (1 mL), dioxane (0.5 mL) and DMF (0.5 mL), undernitrogen atmosphere, was added PyBOP (44.6 mg, 0.09 mmol). Afterstirring at ambient temperature for 15 hours, the mixture was dilutedwith DMSO, passed through a syringe filter, then purified viapreparative HPLC/MS to afford the title compound (21.1 mg; 65% yield).MS(ES): m/z=378 [M+H]⁺. t_(R)=1.25 min (Method I). ¹H NMR (500 MHz,DMSO-d₆) δ 8.63-8.53 (m, 2H), 7.97 (dd, J=8.9, 5.7 Hz, 1H), 7.70 (d,J=7.9 Hz, 2H), 7.63-7.51 (m, 2H), 7.25 (d, J=7.8 Hz, 2H), 7.00 (d, J=4.8Hz, 1H), 3.47 (d, J=11.4 Hz, 2H), 2.76 (t, J=11.6 Hz, 2H), ˜2.53 (m,integration, exact chemical shift range obscured by solvent peak), 2.31(s, 3H), 1.87-1.73 (m, 3H), 1.55-1.41 (m, 2H).

Example 873,4-Dichloro-N-((1-(6-fluoroquinolin-4-yl)piperidin-4-yl)methyl)benzamide

To a heterogeneous mixture of the TFA salt of(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)methanamine (86B, 41.8 mg, 0.09mmol) in anhydrous THF (1 mL) and dioxane (0.5 mL), under nitrogenatmosphere, was added DIPEA (0.06 mL, 0.3 mmol) followed by3,4-dichlorobenzoyl chloride (18.9 mg, 0.09 mmol). After stirring atambient temperature for 16 hours, the mixture was diluted with DMF,filtered through a syringe filter, then purified via preparative HPLC/MSto afford the title compound (23.1 mg; 62% yield). MS(ES): m/z=432[M+H]⁺. t_(R)=1.51 min (Method I). ¹H NMR (500 MHz, DMSO-d₆) δ 8.83-8.76(m, 1H), 8.63 (d, J=4.8 Hz, 1H), 8.07 (s, 1H), 7.99 (dd, J=8.8, 5.7 Hz,1H), 7.82 (d, J=8.2 Hz, 1H), 7.73 (d, J=8.4 Hz, 1H), 7.64-7.54 (m, 2H),7.01 (d, J=4.8 Hz, 1H), 3.71-3.44 (m, 2H), 3.34-3.21 (m, 2H), 2.76 (t,J=11.7 Hz, 2H), 1.88-1.75 (m, 3H), 1.56-1.45 (m, 2H).

Examples 88 to 90

Reaction of the TFA salt of(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)methanamine with an appropriateacid chloride, under the conditions described for Example 87 (Scheme 1,below), afforded compounds of the invention shown in Table 1 below.

TABLE 1 Ex. t_(R) No. R (M + H)⁺ (min.)^(Method) 88

398 1.31¹ 89

382 1.19¹ 90

398 1.33¹

Example 91 1-(1-(6-Fluoroquinolin-4-yl)piperidin-4-yl)-3-(p-tolyl)urea

91A. tert-Butyl (1-(6-fluoroquinolin-4-yl)piperidin-4-yl)carbamate

To a homogeneous mixture of 4-chloro-6-fluoroquinoline (200.0 mg, 1.1mmol) in anhydrous NMP (5 mL), in a sealable vial, was added4-Boc-aminopiperidine (309.0 mg, 1.5 mmol) followed by DIPEA (0.8 mL,4.6 mmol). The vial was sealed and the mixture was stirred at 120° C.for 15 hours. After cooling to room temperature, the reaction mixturewas partitioned between EtOAc and water. The layers were separated andthe aqueous layer was extracted twice more with EtOAc. The organicextracts were combined, washed with brine, dried (Na₂SO₄), filtered andconcentrated in vacuo to afford the crude product which was used withoutfurther purification, based on quantitative yield. MS(ES): m/z=346[M+H]⁺. t_(R)=0.70 min (Method A).

91B. 1-(6-Fluoroquinolin-4-yl)piperidin-4-amine

To a homogeneous mixture of tert-butyl(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)carbamate (380.0 mg, 1.1 mmol)in dioxane (2 mL), under nitrogen atmosphere, was added 4M HCl indioxane (2 mL, 8.0 mmol). The resultant mixture was stirred at ambienttemperature for 6 hours, during which time a precipitate formed. Vacuumfiltration afforded the HCl salt of the title compound as a pale yellowsolid (358 mg, 100% yield) which was used without further purification.MS(ES): m/z=246 [M+H]⁺. t_(R)=0.42 min (Method A).

Example 91 1-(1-(6-Fluoroquinolin-4-yl)piperidin-4-yl)-3-(p-tolyl)urea

To a heterogeneous mixture of the HCl salt of1-(6-fluoroquinolin-4-yl)piperidin-4-amine (91B, 30.0 mg, 0.09 mmol) inanhydrous THF (1 mL), at room temperature, was added DIPEA (0.05 mL,0.29 mmol) followed by 1-isocyanato-4-methylbenzene (15.1 mg, 0.11mmol). The resulting mixture was stirred for 88 hours, before beingdiluted with DMSO, passed through a syringe filter, then purified viapreparative HPLC/MS to afford the title compound (21.7 mg; 60% yield).MS(ES): m/z 379 [M+H]⁺. t_(R)=1.30 min (Method I). ¹H NMR (500 MHz,DMSO-d₆) δ 8.67 (d, J=2.5 Hz, 1H), 8.29 (s, 1H), 8.11-7.96 (m, 1H),7.70-7.57 (m, 2H), 7.27 (d, J=7.3 Hz, 2H), 7.12-6.97 (m, 3H), 6.23 (d,J=7.2 Hz, 1H), 3.79-3.68 (m, 1H), 3.53-3.37 (m, 2H), 3.05-2.90 (m, 2H),2.21 (s, 3H), 2.11-1.98 (m, 2H), 1.81-1.65 (m, 2H).

Example 921-(4-Chloro-2-fluorophenyl)-3-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)urea

Example 92 (19.3 mg; 49% yield) was prepared following a procedureanalogous to that for the synthesis of Example 91, except that4-chloro-2-fluoro-1-isocyanatobenzene (19.4 mg, 0.11 mmol) was usedinstead of 1-isocyanato-4-methylbenzene. MS(ES): m/z=417 [M+H]⁺.T_(r)=1.42 min (Method I). ¹H NMR (500 MHz, DMSO-d₆) δ 8.67 (d, J=4.8Hz, 1H), 8.39 (s, 1H), 8.16 (t, J=8.9 Hz, 1H), 8.02 (dd, J=9.9, 5.6 Hz,1H), 7.68-7.56 (m, 2H), 7.39 (d, J=11.2 Hz, 1H), 7.17 (d, J=8.8 Hz, 1H),7.06 (d, J=4.7 Hz, 1H), 6.84 (d, J=7.3 Hz, 1H), 3.82-3.70 (m, 1H),3.50-3.37 (m, 2H), 2.98 (t, J=10.8 Hz, 2H), 2.12-2.01 (m, 2H), 1.78-1.65(m, 2H).

Example 931-(4-Fluorophenyl)-3-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)urea

Example 93 (21.8 mg; 61% yield) was prepared following a procedureanalogous to that for the synthesis of Example 91, except that1-fluoro-4-isocyanatobenzene (15.5 mg, 0.11 mmol) was used instead of1-isocyanato-4-methylbenzene. MS(ES): m/z=383 [M+H]⁺. T_(r)=1.18 min(Method I). ¹H NMR (500 MHz, DMSO-d₆) δ 8.57 (d, J=6.1 Hz, 1H), 8.44 (s,1H), 8.00 (dd, J=9.2, 5.2 Hz, 1H), 7.84-7.71 (m, 2H), 7.35 (dd, J=8.4,4.9 Hz, 2H), 7.15 (d, J=6.3 Hz, 1H), 7.05 (t, J=8.7 Hz, 2H), 6.31 (d,J=7.5 Hz, 1H), 3.86-3.85 (m, 3H), 3.35 (t, J=11.1 Hz, 2H), 2.07-2.00 (m,2H), 1.74-1.61 (m, 2H).

Example 97 trans-N-(4-Methoxybenzyl)-4-phenylcyclohexanamine

A suspension of trans-4-phenylcyclohexanone (2 g, 11.48 mmol) in DCM (30mL) was treated with (4-methoxyphenyl)methanamine (1.575 g, 11.48 mmol)and magnesium perchlorate (0.128 g, 0.574 mmol). After stirring at rtfor 16 h Na₂SO₄ was added and stirring continued at rt for 1 hr. Themixture was filtered washed with MeOH and the mother liquor was conc. toyield a yellow viscous oil. The oil was dissolved in MeOH (10 mL), andtreated with NaBH₄ (0.651 g, 17.22 mmol) in portions, then stirred at rtfor 30 min until the reaction was done. The reaction was quenched withwater (75 ml) and extracted with EtOAc (3×). The combined organicextracts were dried, filtered, and concentrated. The yellow residue waspurified on an ISCO silica column (40 g), eluting with (DCM:10% MeOH inDCM contain 2.5% NH₄OH=0%-70%) in 20 mins. to yield(1r,4r)-N-(4-methoxybenzyl)-4-phenylcyclohexanamine (870 mg, 2.94 mmol,25%) ¹H NMR (500 MHz, chloroform-d) δ 7.32-7.22 (m, 3H), 7.22-7.10 (m,4H), 6.92-6.77 (m, 2H), 3.84-3.70 (m, 5H), 2.60-2.46 (m, 2H), 2.15-2.02(m, 2H), 1.97-1.82 (m, 2H), 1.49 (qd, J=12.9, 3.0 Hz, 2H), 1.36-1.15 (m,2H) MS: Anal. Calc'd for C₂₀H₂₅NO 295.194. found [M+H] 296.1 LC: tr=1.4min (Method I)

Example 1191-(4-Chlorophenyl)-3-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)urea

119A. trans-4-(6-Fluoroquinolin-4-yl)cyclohexanamine

To a solution of 4-(6-fluoroquinolin-4-yl)cyclohexanone (350 mg, 1.439mmol) in EtOH (6 mL) in a microwave vial was added ammonium acetate(1663 mg, 21.58 mmol). The resulting suspension was treated with sodiumcyanoborohydride (108 mg, 1.726 mmol). The reaction was capped andmicrowaved at 130° C. for 5 min. The reaction was cooled to RT anddiluted with MeOH and purified by preparative HPLC (PHENOMENEX® Luna 5μ30×100 mm), 40 mL/min flow rate with gradient of 0% B-100% B over 12minutes. Hold at 100% B for 2 min. (A: 0.1% TFA in water/MeOH (90:10),B: 0.1% TFA in water/MeOH (10:90) monitoring at 254 nm. Fractionscontaining the product were combined and concentrated to givetrans-4-(6-fluoroquinolin-4-yl)cyclohexanamine (310 mg, 0.624 mmol,43.4% yield)¹H NMR (400 MHz, DMSO-d₆) δ 8.88 (d, J=4.6 Hz, 1H),8.19-8.05 (m, 2H), 7.92 (br. s., 3H), 7.73 (td, J=8.7, 2.8 Hz, 1H), 7.53(d, J=4.6 Hz, 1H), 3.36 (br. s., 1H), 3.22-3.02 (m, 1H), 2.09 (br. s.,2H), 1.96 (br. s., 2H), 1.77-1.52 (m, 4H) MS: Anal. Calc'd for C₁₅H₁₇FN₂244.138. found [M+H] 245.1 LC: t_(r)=0.42 min (Method A)

Example 1191-(4-Chlorophenyl)-3-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)urea

To a solution of trans-4-(6-fluoroquinolin-4-yl)cyclohexanamine (20 mg,0.042 mmol) in THF (0.5 mL) at RT was added 1-chloro-4-isocyanatobenzene(9.76 mg, 0.064 mmol). The reaction was stirred at RT for 3 h. The crudematerial was purified via preparative LC/MS with the followingconditions: Column: XBridge C18, 19×200 mm, 5-μm particles; Mobile PhaseA: 5:95 acetonitrile: water with 10-mM ammonium acetate; Mobile Phase B:95:5 acetonitrile: water with 10-mM ammonium acetate; Gradient: 30-70% Bover 22 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min.Fractions containing the desired product were combined and dried viacentrifugal evaporation. The yield of the product was 11.4 mg (0.029mmol, 67%) ¹H NMR (400 MHz, DMSO-d₆) δ 8.82 (d, J=4.5 Hz, 1H), 8.50 (s,1H), 8.10 (dd, J=9.3, 5.9 Hz, 1H), 8.03 (dd, J=11.1, 2.8 Hz, 1H), 7.68(td, J=8.7, 2.8 Hz, 1H), 7.50 (d, J=4.5 Hz, 1H), 7.46-7.35 (m, 2H),7.33-7.19 (m, 2H), 6.19 (d, J=7.7 Hz, 1H), 3.70-3.52 (m, 1H), 3.42-3.34(m, 1H), 2.04 (d, J=9.3 Hz, 2H), 1.92 (d, J=12.1 Hz, 2H), 1.79-1.63 (m,2H), 1.61-1.45 (m, 2H) MS: Anal. Calc'd for C₂₂H₂₁ClFN₃O 397.136. found[M+H] 398.2 LC: t_(r)=1.39 min (Method I).

These compounds were obtained following the procedures in Example 119using the corresponding isocyanates.

Ex. Tr No. Name R (min)^(Method 1) [M + H]⁺ 1201-(4-chloro-2-fluorophenyl)-3-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)urea

1.46 416.0 121 1-(4-cyanophenyl)-3-trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)urea

1.16 389.1 122 1-(2,4-difluorophenyl)-3-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)urea

1.25 399.9 123 1-(3,4-difluorophenyl)-3-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)urea

1.37 400.2 124 1-(4-fluorophenyl)-3-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)urea

1.26 382.3

Example 1251-(4-Chlorophenyl)-3-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)urea

125A. cis-4-(6-Fluoroquinolin-4-yl)cyclohexanamine

To a solution of 4-(6-fluoroquinolin-4-yl)cyclohexanone (350 mg, 1.439mmol) in EtOH (6 mL) in a microwave vial was added ammonium acetate(1663 mg, 21.58 mmol). To the resulting suspension was added sodiumcyanoborohydride (108 mg, 1.726 mmol). The reaction was capped andmicrowaved at 130° C. for 5 min. The reaction was cooled to RT anddiluted with MeOH and purified by preparative HPLC (PHENOMENEX® Luna 5μ30×100 mm), 40 mL/min flow rate with gradient of 0% B-100% B over 12minutes Hold at 100% B for 2 min. (A: 0.1% TFA in water/MeOH (90:10), B:0.1% TFA in water/MeOH (10:90) monitoring at 254 nm. Fractionscontaining the product were combined and concentrated to givecis-4-(6-fluoroquinolin-4-yl)cyclohexanamine (100 mg, 0.201 mmol, 14%yield)¹H NMR (400 MHz, DMSO-d₆) δ 8.94 (d, J=4.6 Hz, 1H), 8.19-8.03 (m,2H), 7.94 (br. s., 1H), 7.74 (td, J=8.7, 2.8 Hz, 1H), 7.60 (d, J=4.8 Hz,1H), 3.59 (br. s., 1H), 3.49 (t, J=11.0 Hz, 1H), 2.10-1.82 (m, 6H), 1.75(d, J=10.8 Hz, 2H) MS: Anal. Calc'd for C₁₅H₁₇FN₂ 244.138. found [M+H]245.1 LC: t_(r)=0.45 min (Method A).

Example 1251-(4-Chlorophenyl)-3-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)urea

To a solution of cis-4-(6-fluoroquinolin-4-yl)cyclohexanamine (25 mg,0.053 mmol) in THF (0.5 mL) at RT was added 1-chloro-4-isocyanatobenzene(16.27 mg, 0.106 mmol). The reaction was stirred at RT for 2 h. Thecrude material was purified via preparative LC/MS with the followingconditions: Column: XBridge C18, 19×200 mm, 5-μm particles; Mobile PhaseA: 5:95 acetonitrile: water with 10-mM ammonium acetate; Mobile Phase B:95:5 acetonitrile: water with 10-mM ammonium acetate; Gradient: 30-70% Bover 22 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min.Fractions containing the desired product were combined and dried viacentrifugal evaporation. The yield of the product was 14.7 mg (0.034mmol, 64%)¹H NMR (500 MHz, DMSO-d₆) δ 8.95 (d, J=4.5 Hz, 1H), 8.56 (s,1H), 8.21-8.08 (m, 2H), 7.78 (t, J=8.5 Hz, 1H), 7.63 (d, J=4.4 Hz, 1H),7.43 (d, J=8.3 Hz, 2H), 7.27 (d, J=7.4 Hz, 2H), 6.69 (d, J=7.4 Hz, 1H),4.01 (br. s., 1H), 3.59 (d, J=9.4 Hz, 1H), 1.96-1.78 (m, 4H), 1.76 (br.s., 4H) MS: Anal. Calc'd for C₂₂H₂₁ClFN₃O 397.136. found [M+H] 398.2 LC:tr=1.44 min (Method I).

Example 130 trans-N-Benzyl-4-(6-fluoroquinolin-4-yl)cyclohexanamine

To a solution of 4-(6-fluoroquinolin-4-yl)cyclohexanone (100 mg, 0.411mmol) and benyzlamine (66 mg, 0.617 mmol) in CH₂Cl₂ (2 mL) at RT wasadded acetic acid (0.024 mL, 0.411 mmol), followed by sodiumtriacetoxyborohydride (131 mg, 0.617 mmol). The reaction was stirred atRT for 4 h. Then it was diluted with MeOH and purified with prep HPLC(Phen Luna 5u 30×100 mm), 40 mL/min flow rate with gradient of 0% B-100%B over 12 minutes Hold at 100% B for 2 min. (A: 0.1% TFA in water/MeOH(90:10), B: 0.1% TFA in water/MeOH (10:90) monitoring at 254 nm.Evaporation of the product containing fractions gave(1r,4r)-N-benzyl-4-(6-fluoroquinolin-4-yl)cyclohexanamine (150 mg). Analiquot (15 mg) of this material was further purified under thefollowing conditions: Column: XBridge C18, 19×200 mm, 5-μm particles;Mobile Phase A: 5:95 acetonitrile: water with 10-mM ammonium acetate;Mobile Phase B: 95:5 acetonitrile: water with 10-mM ammonium acetate;Gradient: 10-50% B over 18 minutes, then a 5-minute hold at 100% B;Flow: 20 mL/min. Fractions containing the desired product were combinedand dried via centrifugal evaporation. The yield of the product was 3.1mg, ¹H NMR (500 MHz, DMSO-d₆) δ 8.78 (br d, J=4.2 Hz, 1H), 8.08 (br dd,J=8.8, 6.0 Hz, 1H), 7.96 (br d, J=10.6 Hz, 1H), 7.72-7.62 (m, 1H), 7.43(br d, J=7.0 Hz, 3H), 7.37 (br t, J=7.3 Hz, 2H), 7.34-7.25 (m, 1H), 3.94(s, 1H), 3.74-3.63 (m, 1H), 3.29 (br s, 1H), 2.78 (br s, 1H), 2.14 (brs, 2H), 1.99-1.84 (m, 3H), 1.55 (br s, 4H). MS: Anal. Calc'd forC₂₂H₂₃FN₂ 334.185. found [M+H] 335.1 LC: tr=0.78 min (Method I).

Example 131 cis-N-Benzyl-4-(6-fluoroquinolin-4-yl)cyclohexanamine

Example 131 was obtained following the procedures in Example 130 usingthe corresponding cis-4-(6-fluoroquinolin-4-yl)cyclohexanone andbenzylamine. ¹H NMR (500 MHz, DMSO-d₆) δ 8.84 (d, J=3.8 Hz, 1H),8.14-8.04 (m, 1H), 8.00 (d, J=10.8 Hz, 1H), 7.67 (t, J=8.7 Hz, 1H),7.55-7.46 (m, 3H), 7.41 (t, J=7.2 Hz, 2H), 7.35 (d, J=7.2 Hz, 1H), 4.01(br. s., 2H), 3.41 (br. s., 1H), 3.17 (br. s., 1H), 2.09-1.99 (m, 2H),1.99-1.80 (m, 4H), 1.67 (d, J=12.0 Hz, 2H). MS: Anal. Calc'd forC₂₂H₂₃FN₂ 334.185. found [M+H] 335.2 LC: t_(r)=0.90 min (Method I).

Example 1392-(4-(((trans-4-(6-Fluoroquinolin-4-yl)cyclohexyl)amino)methyl)phenyl)aceticacid

Example 139 was obtained following the procedures in Example 130 usingthe corresponding trans-4-(6-fluoroquinolin-4-yl)cyclohexanone and2-(4-(aminomethyl)phenyl)acetic acid hydrochloride ¹H NMR (500 MHz,DMSO-d₆) δ 8.91 (br. s., 2H), 8.83 (d, J=3.9 Hz, 1H), 8.20-8.08 (m, 1H),8.03 (d, J=10.7 Hz, 1H), 7.71 (t, J=8.3 Hz, 1H), 7.48 (d, J=7.9 Hz, 3H),7.35 (d, J=7.3 Hz, 2H), 4.21 (br. s., 2H), 3.60 (d, J=19.4 Hz, 2H),3.41-3.28 (m, 1H), 3.21 (br. s., 1H), 2.28 (d, J=10.6 Hz, 2H), 2.01 (d,J=11.3 Hz, 2H), 1.81-1.59 (m, 4H) MS: Anal. Calc'd for C₂₄H₂₅FN₂O₂392.190. found [M+H] 393.1 LC: tr=0.72 min (Method I).

Example 1402-(4-(((cis-4-(6-Fluoroquinolin-4-yl)cyclohexyl)amino)methyl)phenyl)aceticacid

Example 140 was obtained following the procedures in Example 130 usingthe corresponding cis-4-(6-fluoroquinolin-4-yl)cyclohexanone and2-(4-(aminomethyl)phenyl)acetic acid hydrochloride ¹H NMR (500 MHz,DMSO-d₆) δ 8.81 (d, J=4.2 Hz, 1H), 8.16-8.03 (m, 1H), 7.98 (d, J=9.6 Hz,1H), 7.73-7.60 (m, 1H), 7.51 (d, J=4.3 Hz, 1H), 7.42-7.29 (m, J=7.6 Hz,2H), 7.26-7.08 (m, J=7.5 Hz, 2H), 3.81 (br. s., 1H), 3.64 (br. s., 1H),3.49 (s, 1H), 3.34 (br. s., 1H), 3.16 (s, 1H), 2.99 (br. s., 1H),2.01-1.87 (m, 4H), 1.83-1.68 (m, 2H), 1.61 (d, J=11.9 Hz, 2H). MS: Anal.Calc'd for C₂₄H₂₅FN₂O₂ 392.190. found [M+H] 393.2 LC: tr=0.79 min(Method I).

Example 1442-(4-Chlorophenyl)-N-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)acetamide

To a solution of trans-4-(6-fluoroquinolin-4-yl)cyclohexanamine (20 mg,0.042 mmol) (Intermediate 119A) in THF (0.5 mL) at RT was added2-(4-chlorophenyl)acetyl chloride (16.02 mg, 0.085 mmol), followed bytriethylamine (0.024 mL, 0.169 mmol). The reaction was stirred at RT for3 h. The crude material was purified via preparative LC/MS with thefollowing conditions: Column: XBridge C18, 19×200 mm, 5-μm particles;Mobile Phase A: 5:95 acetonitrile: water with 10-mM ammonium acetate;Mobile Phase B: 95:5 acetonitrile: water with 10-mM ammonium acetate;Gradient: 50-100% B over 20 minutes, then a 5-minute hold at 100% B;Flow: 20 mL/min. Fractions containing the desired product were combinedand dried via centrifugal evaporation. The yield of the product was 11.5mg (0.029 mmol, 68%)¹H NMR (500 MHz, DMSO-d₆) δ 8.76 (d, J=4.5 Hz, 1H),8.25 (d, J=7.7 Hz, 1H), 8.07 (dd, J=9.1, 5.8 Hz, 1H), 7.93 (d, J=8.8 Hz,1H), 7.71-7.58 (m, 1H), 7.46 (d, J=4.4 Hz, 1H), 7.36-7.30 (m, J=8.2 Hz,2H), 7.29-7.13 (m, J=8.2 Hz, 2H), 3.94-3.81 (m, 2H), 3.61 (d, J=7.3 Hz,1H), 3.25 (t, J=11.2 Hz, 1H), 1.88 (t, J=13.7 Hz, 4H), 1.66-1.43 (m, 4H)MS: Anal. Calc'd for C₂₃H₂₂ClFN₂O 396.140. found [M+H] 397.0 LC:t_(r)=1.37 min (Method I).

Ex. Tr No. Name R (min)^(Method 1) [M + H]⁺ 1452-(4-chlorophenyl)-N-(trans-4- (6-fluoroquinolin-4-yl)cyclohexyl)propanamide

1.52 411.1 146 4-chloro-N-(trans-4-(6- fluoroquinolin-4-yl)cyclohexyl)benzamide

1.42 383.2 147 2-(4-chlorophenyl)-N-(cis-4-(6- fluoroquinolin-4-yl)cyclohexyl)acetamide

1.75 397.2 148 2-(4-chlorophenyl)-N-(cis-4-(6- fluoroquinolin-4-yl)cyclohexyl)propanamide

1.47 411.2 149 4-chloro-N-(cis-4-(6- fluoroquinolin-4-yl)cyclohexyl)benzamide

1.30 383.3

These compounds were obtained following the procedures in Example 144using the corresponding amines and acid chloride.

Example 157 a, b, c, d, e4-Chloro-N-(1-(4-(2-methylpyridin-4-yl)cyclohexyl)ethyl)benzamide4-Chloro-N-((S)-1-(cis-4-(2-methylpyridin-4-yl)cyclohexyl)ethyl)benzamide4-Chloro-N-((R)-1-(cis-4-(2-methylpyridin-4-yl)cyclohexyl)ethyl)benzamide4-Chloro-N-((S)-1-(trans-4-(2-methylpyridin-4-yl)cyclohexyl)ethyl)benzamide4-Chloro-N-((R)-1-(trans-4-(2-methylpyridin-4-yl)cyclohexyl)ethyl)benzamide

157A. Ethyl 2-(1,4-dioxaspiro[4.5]decan-8-ylidene)propanoate

To a suspension of NaH (0.307 g, 7.68 mmol) in THF (8 mL) cooled at 0°C. was added ethyl 2-(diethoxyphosphoryl)propanoate (1.830 g, 7.68 mmol)slowly. After 30 min, 1,4-dioxaspiro[4.5]decan-8-one (1 g, 6.40 mmol)was added. The resulting mixture was stirred at 0° C. for 2 h, thenwarmed to RT overnight. The mixture was quenched with water and the THFwas removed in vacuo. The residue was dissolved in EtOAc, washed withwater and brine. The solution was dried over Na₂SO₄, filtered andconcentrated. The crude material was purified by ISCO (EtOAc/Hex 0-30%).Fractions containing the product were concentrated to yield ethyl2-(1,4-dioxaspiro[4.5]decan-8-ylidene)propanoate (1.2 g, 78% yield) alight yellow oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.19 (q, J=7.1 Hz,2H), 4.03-3.89 (m, 4H), 2.68-2.53 (m, 2H), 2.46-2.28 (m, 2H), 1.89 (s,3H), 1.78-1.66 (m, 4H), 1.30 (t, J=7.1 Hz, 3H).

157B. Ethyl 2-(1,4-dioxaspiro[4.5]decan-8-yl)propanoate

A suspension of ethyl 2-(1,4-dioxaspiro[4.5]decan-8-ylidene)propanoate(500 mg, 2.081 mmol) and 10% palladium on carbon (25 mg, 0.024 mmol) inEtOAc (5 mL) was hydrogenated in a Parr shaker at 45 psi for 6 h. Thecatalyst was filtered and the filtrate was concentrated to yield ethyl2-(4-(3-methylpyridin-4-yl)cyclohexyl)propanoate (450 mg, 89% yield) asa light oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.12 (dtt, J=10.7, 7.1,3.6 Hz, 2H), 3.98-3.81 (m, 4H), 2.35-2.17 (m, 1H), 1.83-1.68 (m, 3H),1.66-1.45 (m, 4H), 1.43-1.28 (m, 2H), 1.27-1.22 (m, 3H), 1.14-1.07 (m,3H)

157C. Ethyl 2-(4-oxocyclohexyl)propanoate

To a solution of ethyl 2-(1,4-dioxaspiro[4.5]decan-8-yl)propanoate (450mg, 1.857 mmol) in THF (5 mL) was added 1M hydrogen chloride (aqueous)(0.929 mL, 3.71 mmol). The mixture was heated at 50° C. for 6 h. Thereaction mixture was concentrated. The residue was dissolved in EtOAc,washed with water (2×), and brine. The solution was dried over Na₂SO₄and concentrated. The crude material was purified with ISCO (EtOAc/Hex0-30%). Fractions containing product were concentrated to yield ethyl2-(4-oxocyclohexyl)propanoate (290 mg, 79% yield) as a clear oil. ¹H NMR(400 MHz, CHLOROFORM-d) δ 4.22-4.06 (m, 2H), 2.46-2.30 (m, 5H),2.13-1.91 (m, 3H), 1.56-1.42 (m, 2H), 1.31-1.24 (m, 3H), 1.18 (d, J=7.1Hz, 3H).

157D. Ethyl2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)propanoate

Ethyl 2-(4-oxocyclohexyl)propanoate (200 mg, 1.01 mmol) and2,6-di-tert-butyl-4-methylpyridine (238 mg, 1.16 mmol) were dissolved indry DCM (10 ml). To the reaction mixture trifluoromethanesulfonicanhydride (0.186 mL, 1.11 mmol) was added dropwise and stirred for 2 h.The suspension was filtered and the filtrate was diluted with DCM,washed with 1N HCl (2×), satd. sodium bicarbonate solution, water, andbrine. The solution was dried over Na₂SO₄ and concentrated to yieldethyl 2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)propanoate(320 mg, 96% yield) as a brown oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ5.73 (t, J=6.1 Hz, 1H), 4.28-4.05 (m, 2H), 2.52-2.17 (m, 4H), 2.08-1.79(m, 3H), 1.49 (dt, J=11.1, 6.6 Hz, 1H), 1.31-1.20 (m, 3H), 1.19-1.04 (m,3H)

157E. Ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)propanoate

To a solution of ethyl2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)propanoate (300mg, 0.908 mmol) in DMSO (5 mL) was added4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (230 mg,0.908 mmol) and potassium acetate (267 mg, 2.72 mmol). After the mixturewas degassed with N₂ for 10 minutes, PdCl₂(dppf) (19.9 mg, 0.027 mmol)was added. The mixture was heated at 80° C. overnight. The mixture waspartitioned between EtOAc and water. The organic phase was concentratedand purified by ISCO silica gel column. Fractions containing productwere concentrated to yield ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)propanoate(168 mg, 60% yield) as a brown oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ6.66-6.40 (m, 1H), 4.31-4.00 (m, 2H), 2.34-2.26 (m, 1H), 2.25-2.19 (m,1H), 2.19-2.04 (m, 2H), 1.95-1.75 (m, 3H), 1.73-1.60 (m, 1H), 1.29-1.24(m, 15H), 1.13 (dd, J=11.6, 7.0 Hz, 3H)

157F. Ethyl 2-(4-(2-methylpyridin-4-yl)cyclohex-3-en-1-yl)propanoate

To a solution of ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)propanoate(120 mg, 0.389 mmol) in dioxane (3 mL) was added4-bromo-2-methylpyridine (67.0 mg, 0.389 mmol), water (1 mL) and Na₂CO₃(165 mg, 1.557 mmol). The mixture was degassed with N2 for 10 minutes.Pd(Ph₃P)₄ (22.49 mg, 0.019 mmol) was then added. The mixture was heatedto 100° C. for 16 h. The cooled mixture was diluted with EtOAc, washedwith water and brine. The solution was dried over Na₂SO₄, filtered andevaporated. The crude material was purified by ISCO silica gelchromatography (0-50% EtOAc/Hexane). Fractions containing product wereconcentrated to yield ethyl2-(4-(2-methylpyridin-4-yl)cyclohex-3-en-1-yl)propanoate (100 mg, 0.366mmol, 94% yield) as a yellow oil. ¹H NMR (400 MHz, chloroform-d) δ8.61-8.11 (m, 1H), 7.09-6.68 (m, 2H), 4.15 (qdd, J=7.1, 3.3, 1.8 Hz,2H), 2.71-2.57 (m, 1H), 2.53 (d, J=5.3 Hz, 3H), 2.47-2.35 (m, 0.5H),2.29 (t, J=7.1 Hz, 0.5H), 1.98-1.75 (m, 3H), 1.67-1.38 (m, 4H),1.32-1.22 (m, 4H), 1.21-1.09 (m, 4H).

157G Ethyl 2-(4-(2-methylpyridin-4-yl)cyclohexyl)propanoate

Ethyl 2-(4-(2-methylpyridin-4-yl)cyclohex-3-en-1-yl)propanoate (100 mg,0.366 mmol) was dissolved in MeOH (5 mL). Ammonium formate (115 mg,1.829 mmol) and palladium on carbon (10%) (10.51 mg, 0.099 mmol) wereadded. The vessel was equipped with a reflux condenser, evacuated andflushed with N₂ three times. The reaction was then heated to reflux.After one hour, the reaction was cooled and filtered. The filtrate wasconcentrated in vacuo. The residue was dissolved in EtOAc, washed withsodium bicarbonate solution, water, and brine. The solution was driedover Na₂SO₄, filtered and concentrated. The crude product was useddirectly in the next step.

157H. 2-(4-(2-Methylpyridin-4-yl)cyclohexyl)propanoic acid

To a mixture of ethyl 2-(4-(2-methylpyridin-4-yl)cyclohexyl)propanoate(320 mg, 1.162 mmol) in THF (2 mL), MeOH (2 mL) and water was added LiOH(278 mg, 11.62 mmol). The mixture was heated at 70° C. for 4 h. LC-MSindicated the completion of the reaction. The mixture was cooled to RT,neutralized with 1N HCl until pH-4, and extracted with EtOAc 3 times.The combined organic phases were washed with water and brine. Thesolution was dried over Na₂SO₄ and concentrated. ¹H NMR (400 MHz,chloroform-d) δ 8.41 (d, J=5.3 Hz, 1H), 7.09-6.94 (m, 2H), 2.75-2.59 (m,1H), 2.54 (d, J=3.5 Hz, 3H), 2.44 (br. s., 1H), 2.35 (t, J=7.0 Hz, 1H),1.98-1.82 (m, 3H), 1.77-1.61 (m, 4H), 1.56-1.39 (m, 1H), 1.20 (d, J=6.7Hz, 3H); MS: Anal. Calc'd for C₁₅H₂₁NO₂ 247.16. found [M+H] 248.08 LC:t_(r)=0.55 min.

157I. 1-(4-(2-Methylpyridin-4-yl)cyclohexyl)ethanamine

2-(4-(2-Methylpyridin-4-yl)cyclohexyl)propanoic acid (240 mg, 0.970mmol) (157B) was taken up in toluene (5 ml) and diphenyl phosphorazidate(0.230 mL, 1.067 mmol) and triethylamine (0.162 mL, 1.164 mmol) wereadded. The vial was sealed and heated to 70° C. After about 2 h, thereaction was cooled to rt and concentrated under reduced pressure. Thecrude residue was taken up in 40 mL THF and 40 mL of water and lithiumhydroxide (1.589 g, 66.4 mmol) was added. The reaction was stirred atrt. LCMS after 1 hour shows that the isocyanate was consumed. Thereaction was acidified with 1N HCl (white precipitate forms) andextracted with EtOAc to remove DPPA related impurities. The solution wasmade basic with 1N NaOH (precipitate forms again) and extracted withEtOAc (×5). The basic extracts were concentrated in vacuo to give1-(4-(2-methylpyridin-4-yl)cyclohexyl)ethanamine (140 mg, 0.641 mmol,66.1% yield) as a yellow oil. ¹H NMR (400 MHz, chloroform-d) δ 8.65-8.17(m, 1H), 7.08-6.86 (m, 2H), 2.97 (dd, J=8.6, 6.4 Hz, 0.5H), 2.79-2.62(m, 1H), 2.52 (d, J=2.8 Hz, 3H), 2.48-2.33 (m, 0.5H), 2.03-1.90 (m, 2H),1.90-1.68 (m, 4H), 1.50-1.11 (m, 3H), 1.08 (dd, J=6.4, 2.8 Hz, 3H). MS:Anal. Calc'd for C₁₄H₂₂N₂ 218.18. found [M+H] 219.2 LC: t_(r)=0.43 min.

Example 157a4-Chloro-N-(1-(4-(2-methylpyridin-4-yl)cyclohexyl)ethyl)benzamide

To a solution of 1-(4-(2-methylpyridin-4-yl)cyclohexyl)ethanamine (100mg, 0.458 mmol) (157I) in THF (2 mL) was added 4-chlorobenzoic acid (108mg, 0.687 mmol), HOBT (140 mg, 0.916 mmol), EDC (176 mg, 0.916 mmol) andTEA (0.192 mL, 1.374 mmol). The mixture was stirred at RT overnight. Thereaction mixture was filtered and purified via preparative LC/MS withthe following conditions: Column: XBridge C18, 19×200 mm, 5-μmparticles; Mobile Phase A: 5:95 acetonitrile: water with 0.1%trifluoroacetic acid; Mobile Phase B: 95:5 acetonitrile: water with 0.1%trifluoroacetic acid; Gradient: 25-100% B over 20 minutes, then a4-minute hold at 100% B; Flow: 20 mL/min. Fractions containing thedesired product were combined and dried via centrifugal evaporation toyield 4-chloro-N-(1-(4-(2-methylpyridin-4-yl)cyclohexyl)ethyl)benzamide(136.8 mg, 84% yield). ¹H NMR (500 MHz, DMSO-d₆) δ 8.52-8.20 (m, 2H),7.84 (dd, J=10.6, 8.7 Hz, 2H), 7.52 (dd, J=8.4, 2.4 Hz, 2H), 7.27-6.85(m, 2H), 4.24 (br. s., 0.5H), 3.87 (d, J=7.4 Hz, 0.5H), 3.62-3.37 (m,1H), 2.42 (d, J=9.1 Hz, 3H), 1.94-1.31 (m, 8H), 1.20-1.02 (m, 4H).

Example 157b, c, d, e4-Chloro-N-(1-(4-(2-methylpyridin-4-yl)cyclohexyl)ethyl)benzamide(Homochiral with Absolute and Relative Stereochemistry not Determined)

The material was further purified through chiral separation. Anapproximately 140 mg sample was resolved. The material was purified viapreparative SFC with the following conditions: Column: Chiral AD 25×3 cmID, 5-μm particles; Mobile Phase: 70/30 CO₂/MeOH; Detector Wavelength:220 nm; Flow: 85 mL/min. The fractions (“Peak-1” t_(r)=10.117, “Peak-2”t_(r)=11.355, “Peak-3” t_(r)=14.873 and “Peak-4” t_(r)=18.312;analytical conditions: Column: Chiral AD 250×4.6 mm ID, 5 μm particles;Mobile Phase: 70/30 CO₂/MeOH Flow: 2.0 mL/min) were collected in MeOH.

157b First eluting isomer: ¹H NMR (500 MHz, DMSO-d₆) δ 8.44-8.18 (m,2H), 7.84 (d, J=8.3 Hz, 2H), 7.52 (d, J=8.3 Hz, 2H), 7.24-6.96 (m, 2H),4.26 (d, J=6.9 Hz, 1H), 2.60 (br. s., 1H), 2.43 (s, 3H), 1.84-1.36 (m,9H), 1.14 (d, J=6.5 Hz, 3H). MS: Anal. Calc'd for C₂₁H₂₅ClN₂O 356.17.found [M+H] 357.0 LC: t_(r)=1.826 (Method A).

157c Second eluting isomer: ¹H NMR (500 MHz, DMSO-d₆) δ 8.48-8.19 (m,2H), 7.81 (d, J=8.3 Hz, 2H), 7.50 (d, J=8.3 Hz, 2H), 7.27-6.82 (m, 2H),4.24 (d, J=6.9 Hz, 1H), 2.60 (br. s., 1H), 2.42 (s, 3H), 1.83-1.37 (m,9H), 1.13 (d, J=6.4 Hz, 3H). MS: Anal. Calc'd for C₂₁H₂₅ClN₂O 356.17.found [M+H] 356.9 LC: t, =1.864 (Method A).

157d Third eluting isomer: ¹H NMR (500 MHz, DMSO-d₆) δ 8.28 (d, J=5.6Hz, 2H), 7.86 (d, J=8.4 Hz, 2H), 7.52 (d, J=8.4 Hz, 2H), 7.23-6.87 (m,2H), 3.87 (d, J=6.4 Hz, 1H), 2.40 (s, 4H), 1.96-1.71 (m, 4H), 1.58-1.28(m, 3H), 1.21-0.99 (m, 5H). MS: Anal. Calc'd for C₂₁H₂₅ClN₂O 356.17.found [M+H] 356.9 LC: t_(r)=1.857 (Method A).

157e Fourth eluting isomer: ¹H NMR (500 MHz, DMSO-d₆) δ 8.28 (d, J=4.9Hz, 2H), 7.86 (d, J=8.3 Hz, 2H), 7.52 (d, J=8.3 Hz, 2H), 7.24-6.84 (m,2H), 4.06-3.74 (m, 1H), 2.46-2.28 (m, 4H), 1.95-1.71 (m, 4H), 1.61-1.29(m, 3H), 1.21-1.02 (m, 5H) MS: Anal. Calc'd for C₂₁H₂₅ClN₂O 356.17.found [M+H] 356.8 LC: t_(r)=1.857 (Method A).

The following compounds were obtained using the procedures in Example157.

Ex. Tr No. Name R (min)^(Method) [M + H]⁺ Stereochemistry 158a4-chloro-N-(1-(4-(2-fluoro-3- methylpyridin-4-yl)cyclohexyl)ethyl)benzamide

 2.075^(A) 375.2 Diastereomer Mixture 158b 4-chloro-N-(1-(4-(2-fluoro-3-methylpyridin-4- yl)cyclohexyl)ethyl)benzamide

14.485^(W) 375.1 Homochiral with absolute and relative stereochemistrynot determined 158c 4-chloro-N-(1-(4-(2-fluoro-3- methylpyridin-4-yl)cyclohexyl)ethyl)benzamide

17.193^(W) 375.2 Homochiral with absolute and relative stereochemistrynot determined 158d 4-chloro-N-(1-(4-(2-fluoro-3- methylpyridin-4-yl)cyclohexyl)ethyl)benzamide

19.497^(W) 375.2 Homochiral with absolute and relative stereochemistrynot determined 158e 4-chloro-N-(1-(4-(2-fluoro-3- methylpyridin-4-yl)cycloheyxl)ethyl)benzamide

21.901^(W) 375.1 Homochiral with absolute and relative stereochemistrynot determined) 159a 4-chloro-N-(1-(4-(2,3- dimethylpyridin-4-yl)cyclohexyl)ethyl)benzamide

 1.899^(A) 370.9 Diastereomer Mixture 159b 4-chloro-N-(1-(4-(2,3-dimethylpyridin-4- yl)cyclohexyl)ethyl)benzamide

 7.917^(X) 371.3 Homochiral with absolute and relative stereochemistrynot determined) 159c 4-chloro-N-(1-(4-(2,3- dimethylpyridin-4-yl)cyclohexyl)ethyl)benxamide

 8.920^(X) 371.2 Homochiral with absolute and relative stereochemistrynot determined) 159d 4-chloro-N-(1-(4-(2,3- dimethylpyridin-4-yl)cyclohexyl)ethyl)benzamide

10.505^(X) 371.2 Homochiral with absolute and relative stereochemistrynot determined) 159e 4-chloro-N-(1-(4-(2,3- dimethylpyridin-4-yl)cyclohexyl)ethyl)benzamide

11.426^(X) 371.2 Homochiral with absolute and relative sterochemistrynot determined) 160a 4-chloro-N-(1-(4-(3- methylpyridin-4-yl)cyclohexyl)ethyl)benzamide

 1.912^(A) 357.2 Diastereomer Mixture 160b 4-chloro-N-(1-(4-(3-methylpyridin-4- yl)cyclohexyl)ethyl)benzamide

10.662^(O) 357.3 Homochiral with absolute and relative stereochemistrynot determined) 160c 4-chloro-N-(1-(4-(3- methylpyridin-4-yl)cyclohexyl)ethyl)benzamide

13.158^(Y) 357.2 Homochiral with absolute and relative stereochemistrynot deterermined) 160d 4-chloro-N-(1-(4-(3- methylpyridin-4-yl)cyclohexyl)ethyl)benzamide

14.889^(Y) 357.2 Homochiral with absolute and relative stereochemistrynot determined) 160e 4-chloro-N-(1-(4-(3- methylpyridin-4-yl)cyclohexyl)ethyl)benzamide

19.795^(Y) 357.3 Homochiral with absolute and relative stereochemistrynot determined) 161a 4-choro-N-(1-(4-(3- fluoropyridin-4-yl)cyclohexyl)ethyl)benzamide

 7.542^(Z) 361.2 Homochiral with absolute and relative stereochemistrynot determined) 161b 4-chloro-N-(1-(4-_(3- fluoropyridin-4-yl)cyclohexyl)ethyl)benzamide

 8.044^(Z) 361.4 Homochiral with absolute and relative stereochemistrynot determined) 161c 4-chloro-N-(1-(4-(3- fluoropyridin-4-yl)cyclohexyl)ethyl)benzamide

10.057^(Z) 361.2 Homochiral with absolute and relative stereochemistrynot determined) 161d 4-chloro-N-(1-(4-(3- fluoropyridin-4-yl)cyclohexyl)ethyl)benzamide

11.177^(Z) 361.3 Homochiral with absolute and relative sterochemistrynot determined)

Example 1635-Ethoxy-N-((R)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)picolinamide

163A. Methyl 5-ethoxypicolinate

To a solution of methyl 5-hydroxypicolinate (0.1 g, 0.653 mmol) in DMF(2 mL) were added EtI (0.06 mL, 0.72 mmol), and K₂CO₃ (0.135 g, 0.980mmol). The reaction mixture was stirred at rt for 2 h. The reactionmixture was diluted with saturated NaHCO₃ solution and ethyl acetate.The organic layer was separated and concentrated in vacuo to giveIntermediate 163A (white solid, 0.09 g, 0.497 mmol, 76% yield). LC-MSAnal. Calc'd for C₉H₁₁NO₃ 181.07. found [M+H] 182.1, T_(r)=0.66 min(Method A). ¹H NMR (400 MHz, methanol-d₄) δ: 8.29 (d, J=2.6 Hz, 1H),8.11 (dd, J=8.6, 0.4 Hz, 1H), 7.48 (dd, J=8.7, 3.0 Hz, 1H), 4.20 (q,J=7.0 Hz, 2H), 3.94 (s, 3H), 1.45 (t, J=6.9 Hz, 3H).

163B. 5-Ethoxypicolinic acid

To a solution of methyl 5-ethoxypicolinate (0.09 g, 0.497 mmol) in THF(1 mL) and MeOH (1 mL) was added lithium hydroxide solution (1.49 mL,2.98 mmol). The reaction mixture was stirred at rt for 3 h. The reactionmixture was diluted with 1 N HCl solution and ethyl acetate. The organiclayer was separated and dried over MgSO₄. The filtrate was concentratedin vacuo to give Intermediate 163B (white solid, 0.06 g, 0.359 mmol,72.3% yield). LC-MS Anal. Calc'd for C₈H₉NO₃ 167.06. found [M+H] 168.1,T_(r)=0.49 min (Method A). ¹H NMR (400 MHz, DMSO-d₆) δ: 12.75 (br. s.,1H), 8.35 (d, J=2.6 Hz, 1H), 8.01 (d, J=8.6 Hz, 1H), 7.48 (dd, J=8.8,2.9 Hz, 1H), 4.19 (q, J=6.9 Hz, 2H), 1.37 (t, J=6.9 Hz, 3H)

Example 1635-Ethoxy-N-((R)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)picolinamide

To a solution of 5-ethoxypicolinic acid (14.36 mg, 0.086 mmol) in DMF (1mL) was added HATU (33 mg, 0.086 mmol). The reaction mixture was stirredat rt for 5 min, followed by addition of(R)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethanamine (18 mg, 0.066mmol) Intermediate 40L and N-methylmorpholine (0.032 mL, 0.264 mmol).The resulting mixture was stirred at rt for 2 h. The reaction mixturewas concentrated in vacuo and the residue was dissolved in MeOH,filtered, and purified via preparative HPLC to give Example 163 (16 mg,0.038 mmol, 57% yield). LC-MS Anal. Calc'd for C₂₅H₂₈FN₃O₂ 421.22. found[M+H] 422.3. T_(r)=1.63 min (Method I). ¹H NMR (500 MHz, DMSO-d₆) δ:8.81 (d, J=4.4 Hz, 1H), 8.36 (d, J=9.6 Hz, 1H), 8.26 (d, J=2.4 Hz, 1H),8.07 (dd, J=9.1, 5.8 Hz, 1H), 7.99-7.85 (m, 2H), 7.73-7.59 (m, 1H),7.55-7.39 (m, 2H), 4.39 (d, J=6.6 Hz, 1H), 4.14 (q, J=6.9 Hz, 2H),3.71-3.52 (m, 1H), 1.94-1.52 (m, 9H), 1.34 (t, J=6.9 Hz, 3H), 1.19 (d,J=6.4 Hz, 3H).

Example 164a, b, c, d4-Chloro-N-((R)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide4-Chloro-N-((S)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide4-Chloro-N-((R)-1-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide4-Chloro-N-((S)-1-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide(Homochiral with Absolute and Relative Stereochemistry not Determined)

164A. Ethyl 2-(1,4-dioxaspiro[4.5]decan-8-ylidene)acetate

To the flask containing sodium hydride (46.1 g, 1153 mmol) was added THF(1200 mL) at 0° C. under nitrogen. Then triethyl phosphonoacetate (258g, 1153 mmol) was added dropwise. The reaction mixture was stirred at 0°C. for 30 minutes. Then 1,4-dioxaspiro[4.5]decan-8-one (150 g, 960 mmol)was added and stirred at 0° C. for 2 h. The reaction mixture was warmedto rt and stirred for 16 h. The reaction was quenched with water (500mL) and the mixture was concentrated in vacuo. The residue was extractedwith ethyl acetate (3×1000 mL). The combined, organic layers were washedwith water (500 mL) and brine (500 mL) successively. The filtrate wasdried over sodium sulfate and concentrated in vacuo. The crude materialwas purified through flash column chromatography, eluting with 0-30%ethyl acetate in petroleum ether to give Intermediate 164A (pale yellowoil, 135 g, 597 mmol, 62.1% yield). LC-MS Anal. Calc'd for C₁₂H₁₈O₄,226.12 found [M+H]. ¹H NMR (400 MHz, chloroform-d) δ: 5.66 (s, 1H), 4.14(q, J=7.2 Hz, 2H), 4.02-3.82 (m, 4H), 3.24-2.86 (m, 2H), 2.63-2.27 (m,2H), 1.98-1.68 (m, 4H), 1.27 (t, J=7.2 Hz, 3H).

164B. Ethyl 2-(1,4-dioxaspiro[4.5]decan-8-yl)acetate

Ethyl 2-(1,4-dioxaspiro[4.5]decan-8-ylidene)acetate (13.88 g, 61.3 mmol)was taken up in EtOAc (61.3 ml) and was added to a Parr hydrogenationbottle containing 10% palladium on carbon (1.306 g, 12.27 mmol) (54% w/wwater) under an atmosphere of nitrogen. The reaction bottle was purgedwith nitrogen, then with hydrogen. After filling the bottle withhydrogen to 50 psi, the bottle was placed in a Parr shaker and shaken.After 4 hours, the reaction mixture was filtered over pressed CELITE®and concentrated in vacuo to give Intermediate 164B ethyl2-(1,4-dioxaspiro[4.5]decan-8-yl)acetate (colorless oil, 13.78 g, 60.4mmol, 98% yield). LC-MS Anal. Calc'd for C₁₂H₂₀O₄ 228.14. found [M+H]229.1. T_(r)=0.83 min (Method A). ¹H NMR (400 MHz, chloroform-d) δ:4.31-4.08 (m, 2H), 4.00-3.86 (m, 4H), 2.22 (d, J=7.0 Hz, 2H), 1.91-1.79(m, 1H), 1.78-1.70 (m, 4H), 1.63-1.50 (m, 2H), 1.37-1.14 (m, 5H).

164C. Ethyl 2-(4-oxocyclohexyl)acetate

In a 10 liter reactor was taken ethyl2-(1,4-dioxaspiro[4.5]decan-8-yl)acetate (67.5 g, 296 mmol) in acetone(5000 mL). To the reaction mixture was added 1 M HCl solution (1183 mL,1183 mmol) and the resulting mixture was heated under reflux for 2 h.The reaction mixture was concentrated to remove acetone. The residue wasextracted with ethyl acetate (3×1000 mL). Combined organic layer waswashed with water and brine. The organic layer was dried over sodiumsulfate and concentrated in vacuo. The crude material was purifiedthrough flash column chromatography, eluting with 0-20% ethyl acetate inpetroleum ether to give Intermediate 164C (pale yellow liquid, 40 g, 217mmol, 73.4% yield). GC-MS Anal. Calc'd for C₁₀H₁₆O₃, 184.11 found [M]184. T_(r)=10.03 min (Method J).

164D. Ethyl 2-(4-(trifluoromethylsulfonyloxy)cyclohex-3-enyl)acetate

A 2 liter 4 neck flask was charged with2,6-di-tert-butyl-4-methylpyridine (84 g, 407 mmol) in dichloromethane(500 mL) under nitrogen. Tf₂O (55.0 mL, 326 mmol) was added dropwise.Then a solution of ethyl 2-(4-oxocyclohexyl)acetate (50 g, 271 mmol) indichloromethane (500 mL) was added slowly. After completion of theaddition, the reaction mixture was stirred at rt overnight. The reactionmixture was diluted with 1000 mL of dichloromethane and washed withwater and sodium carbonate solution and then water. The organic layerwas dried over sodium sulfate and concentrated in vacuo. The crudematerial was purified through flash column chromatography, eluting with0-10% ethyl acetate in petroleum ether to give Intermediate 164D (paleyellow oil, 65 g, 206 mmol, 76% yield). GC-MS Anal. Calc'd forC₁₁H₁₅F₃O₅S, 316.06 found [M] 317. T_(r)=10.16 min (Method J).

164E. Ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)acetate

In 2 liter 4 neck flask was taken ethyl2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)acetate (120 g,379 mmol), BISPIN (106 g, 417 mmol), and potassium acetate (112 g, 1138mmol) in 1,4-dioxane (1200 mL) under nitrogen. Nitrogen was purgedinside the reaction mixture for 10 minutes. Then1,1′-bis(diphenylphosphino)ferrocene-palladium dichloridedichloromethane complex (15.49 g, 18.97 mmol) was added. The reactionmixture was heated at 80° C. for 16 h. The reaction mixture wasconcentrated. The residue was partitioned between ethyl acetate andwater, filtered through CELITE® bed. The organic layer was separated andthe aqueous layer was extracted with ethyl acetate (3×). Combinedorganic layer was washed with water, brine, and dried over sodiumsulfate and concentrated in vacuo. The crude material was purifiedthrough flash column chromatography, eluting with 0-10% ethyl acetate inpetroleum ether to give Intermediate 164E (pale yellow oil, 56 g, 190mmol, 50.2% yield). GC-MS Anal. Calc'd for C₁₆H₂₇BO₄, 294.20 found [M]295.3. T_(r)=1.10 min (Method A). ¹H NMR (400 MHz, chloroform-d) δ: 6.52(dd, J=4.1, 1.9 Hz, 1H), 4.14 (q, J=7.1 Hz, 2H), 2.62-1.97 (m, 6H),1.94-1.68 (m, 2H), 1.33-1.21 (m, 16H).

164F. Ethyl 2-(4-(6-fluoroquinolin-4-yl)cyclohex-3-en-1-yl)acetate

Ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)acetate(Intermediate 164E) (5 g, 17.00 mmol) was taken up in dioxane (28.3 ml)and water (7.08 ml). 4-Chloro-6-fluoroquinoline (2.57 g, 14.15 mmol) wasadded followed by K₂CO₃ (5.87 g, 42.5 mmol). Mixture was bubbled withnitrogen gas for 5 minutes before the addition of Pd(Ph₃P)₄ (0.327 g,0.283 mmol). After the addition, the reaction was evacuated andbackfilled with N₂ three times and then sealed (sealed vial parafilmed)and heated to 100° C. for 16 hours. The reaction was concentrated invacuo and purified directly via silica gel flash column chromatographyto give Intermediate 164F (4.22 g, 13.47 mmol, 95% yield). LC-MS Anal.Calc'd for C₁₉H₂₀FNO₂ 313.15. found [M+H] 314.1 T_(r)=0.75 min (MethodA).

164G. Ethyl 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)acetate

Intermediate 164F (4.22 g, 13.47 mmol) was dissolved in MeOH (67.3 ml)and ammonium formate (4.25 g, 67.3 mmol) was added. The vessel wasequipped with a reflux condenser and evacuated and flushed with nitrogengas 3 times. Then palladium on carbon (0.143 g, 1.347 mmol) (wet,Degussa type) was added and the reaction was heated to reflux for 1hour. The reaction was cooled, concentrated in vacuo, and diluted withDCM. Solids were filtered off and the filtrate was concentrated to givecrude Intermediate 164G (4.20 g, 13.32 mmol, 99% yield) as a mixture ofcis- and trans-diastereomers. LC-MS Anal. Calc'd for C₁₉H₂₂FNO₂ 315.16.found [M+H] 316.2 T_(r)=0.76 min (Method A).

164H. Ethyl 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)butanoate

To the flask containing THF (6 mL) was added lithium diisopropylamide(2.0 M solution in THF) (3.17 mL, 6.34 mmol) at −78° C., followed byaddition of 1,3-dimethyltetrahydropyrimidin-2(1H)-one (0.573 mL, 4.76mmol) and a solution of ethyl2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)acetate (1.0 g, 3.17 mmol) in THF(10 mL) dropwise at −78° C. The resulting mixture turned into a brownsolution and was stirred at −78° C. for 1 h, then iodoethane (0.51 mL,6.34 mmol) was added slowly. The reaction mixture was then stirred atice bath temperature for 1 h, warmed to rt overnight. The reaction wasquenched by pouring into water and extracting with EtOAc. The combinedorganic layers were washed with brine, dried with MgSO₄, filtered andconcentrated in vacuo. The residue was dissolved in DCM and purified bysilica gel flash chromatography, eluting with 0-20% ethyl acetate inhexane to give Intermediate 164H (oil, 0.81 g, 2.359 mmol, 74.4% yield).LC-MS Anal. Calc'd for C₂₁H₂₆FNO₂, 343.19 found [M+H] 344.3.T_(r)=0.87-0.88 min (Method A). ¹H NMR (400 MHz, chloroform-d) δ:8.88-8.77 (m, 1H), 8.18-8.06 (m, 1H), 7.66 (dd, J=10.6, 2.6 Hz, 1H),7.47 (ddd, J=9.2, 8.0, 2.9 Hz, 1H), 7.36 (d, J=4.6 Hz, 1H), 4.25-4.15(m, 2H), 3.34-3.09 (m, 1H), 2.70-2.16 (m, 1H), 2.13-1.49 (m, 13H),1.36-1.24 (m, 3H), 1.00-0.90 (m, 3H).

164I. 2-(4-(6-Fluoroquinolin-4-yl)cyclohexyl)butanoic acid

To a solution of ethyl 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)butanoate(0.81 g, 2.359 mmol) in THF (4 mL) and MeOH (7 mL) was added 2.0 M LiOHsolution (7.1 mL, 14.2 mmol) slowly. The reaction mixture was stirred atrt overnight. The next day, more LiOH solution (7.1 mL, 14.2 mmol) wasadded to the reaction and the resulting mixture was heated at 70° C. for28 h. The reaction mixture was cooled and ethyl acetate was added. Theaqueous layer was separated and to the aqueous layer was added 1N HClsolution to adjust pH to 5-6. The resulting mixture was diluted withwater and CHCl₃: 2-propanol (2:1). The organic layer was separated anddried over MgSO₄. The filtrate was concentrated in vacuo to giveIntermediate 164I as a mixture of cis- and trans-(3:2) isomers (0.64 g,2.029 mmol, 86% yield). LC-MS Anal. Calc'd for C₁₉H₂₂FNO₂ 315.16 found[M+H] 316.3. T_(r)=0.72 min (Method A). ¹H NMR (400 MHz, chloroform-d)δ: 8.83 (d, J=4.4 Hz, 1H), 8.30-8.03 (m, 1H), 7.67 (dd, J=10.6, 2.4 Hz,1H), 7.48 (ddd, J=9.2, 7.9, 2.6 Hz, 1H), 7.38 (d, J=4.6 Hz, 1H),7.32-7.27 (m, 1H), 3.37-3.07 (m, 1H), 2.77-2.21 (m, 1H), 2.11-1.30 (m,11H), 1.07-1.00 (m, 3H).

164J. 1-(4-(6-Fluoroquinolin-4-yl)cyclohexyl)propan-1-amine

To a suspension of 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)butanoic acid(0.31 g, 0.983 mmol) in toluene (8 mL) were added diphenylphosphorylazide (0.245 mL, 1.13 mmol) and triethylamine (0.15 mL, 1.28 mmol). Thereaction mixture turned into clear solution after addition of TEA. Thevial was sealed and heated to 70° C. for 2.5 h. The reaction mixture wasconcentrated under reduced pressure. To the residue was added THF (10mL) and 2.0 M lithium hydroxide solution (4.91 mL, 9.83 mmol) and theresulting mixture was stirred at rt for 1 h. The reaction mixture wasacidified with 1N HCl (white precipitate forms) and extracted with EtOActo remove DPPA related impurities. Then the aqueous layer was basifiedwith 1N NaOH (precipitate forms again) and extracted with EtOAc fourtimes. The basic extracts were combined, dried over MgSO₄ and thefiltrate was concentrated in vacuo to give colorless oil as a mixture ofcis- and trans-, dried on high vacuum over night to give Intermediate164J (oil, 0.245 g, 0.855 mmol, 87% yield). LC-MS Anal. Calc'd forC₁₈H₂₃FN₂ 286.19. found [M+H] 287.3. T_(r)=0.54 min, 0.55 min (MethodA). ¹H NMR (400 MHz, chloroform-d) δ: 8.81 (d, J=4.6 Hz, 1H), 8.12 (dd,J=9.1, 5.8 Hz, 1H), 7.67 (dd, J=10.6, 2.6 Hz, 1H), 7.47 (ddd, J=9.2,8.0, 2.9 Hz, 1H), 7.37-7.28 (m, 1H), 3.41-3.09 (m, 1H), 2.97-2.50 (m,1H), 2.19-1.23 (m, 11H), 1.06-0.93 (m, 3H).

Example 1644-Chloro-N-(1-(4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide

To a solution of 4-chlorobenzoic acid (42.6 mg, 0.272 mmol) in DMF (2mL) was added HATU (104 mg, 0.272 mmol). The reaction mixture wasstirred at rt for 10 min, followed by addition of1-(4-(6-fluoroquinolin-4-yl)cyclohexyl)propan-1-amine (60 mg, 0.210mmol) (Intermediate 164J) in THF (0.5 mL) and N-methyl morpholine (0.10mL, 0.838 mmol). The resulting mixture was stirred at rt for 2 h. Thereaction mixture was concentrated in vacuo and the residue was dissolvedin MeOH, filtered, and purified via preparative HPLC to give a mixturecontaining the four isomers. The isomers were further separated bypreparative SFC (Method C) to give:

First eluting Example 164a (15 mg, 0.035 mmol, 16.7% yield). LC-MS Anal.Calc'd for C₂₅H₂₆ClFN₂O 424.17 found [M+H] 424.9 T_(r)=1.57 min (MethodI). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.82 (d, J=3.9 Hz, 1H), 8.19 (d, J=9.1Hz, 1H), 8.12-8.03 (m, 1H), 7.94 (d, J=10.3 Hz, 1H), 7.86 (d, J=8.0 Hz,2H), 7.65 (t, J=7.9 Hz, 1H), 7.52 (d, J=8.0 Hz, 2H), 7.46 (d, J=3.5 Hz,1H), 4.27 (d, J=8.0 Hz, 1H), 3.37 (br. s., 1H), 1.92-1.54 (m, 10H), 1.40(d, J=6.2 Hz, 1H), 0.86 (t, J=6.9 Hz, 3H).

Second eluting Example 164b (8.6 mg, 0.020 mmol, 9.6% yield). LC-MSAnal. Calc'd for C₂₅H₂₆ClFN₂O 424.17 found [M+H] 424.9 T_(r)=1.55 min(Method I). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.78 (d, J=4.5 Hz, 1H), 8.17(d, J=9.0 Hz, 1H), 8.07 (dd, J=9.0, 5.8 Hz, 1H), 7.97 (d, J=9.0 Hz, 1H),7.90 (d, J=8.3 Hz, 2H), 7.71-7.59 (m, 1H), 7.54 (d, J=8.3 Hz, 2H), 7.43(d, J=4.4 Hz, 1H), 3.80 (br. s., 1H), 3.27 (t, J=11.3 Hz, 1H), 1.97-1.81(m, 4H), 1.74-1.29 (m, 7H), 0.86 (t, J=7.2 Hz, 3H).

Third eluting Example 164c (6.5 mg, 0.015 mmol, 6.9% yield). LC-MS Anal.Calc'd for C₂₅H₂₆ClFN₂O 424.17 found [M+H] 425.0 T_(r)=1.55 min (MethodI). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.78 (d, J=4.4 Hz, 1H), 8.17 (d, J=9.1Hz, 1H), 8.07 (dd, J=8.9, 5.8 Hz, 1H), 7.97 (d, J=8.9 Hz, 1H), 7.90 (d,J=8.3 Hz, 2H), 7.71-7.60 (m, 1H), 7.54 (d, J=8.3 Hz, 2H), 7.43 (d, J=4.3Hz, 1H), 3.81 (br. s., 1H), 3.36-3.21 (m, 1H), 1.90 (d, J=12.5 Hz, 4H),1.73-1.28 (m, 7H), 0.86 (t, J=7.1 Hz, 3H).

Fourth eluting Example 164d (13.9 mg, 0.032 mmol, 15.5% yield). LC-MSAnal. Calc'd for C₂₅H₂₆ClFN₂O 424.17 found [M+H] 425.1 T_(r)=1.58 min(Method I). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.82 (d, J=4.3 Hz, 1H), 8.19(d, J=9.1 Hz, 1H), 8.08 (dd, J=9.0, 5.9 Hz, 1H), 7.95 (d, J=9.5 Hz, 1H),7.87 (d, J=8.3 Hz, 2H), 7.65 (t, J=7.4 Hz, 1H), 7.53 (d, J=8.3 Hz, 2H),7.46 (d, J=4.3 Hz, 1H), 4.28 (d, J=7.8 Hz, 1H), 3.37 (br. s., 1H),1.92-1.53 (m, 10H), 1.39 (dt, J=14.7, 7.6 Hz, 1H), 0.87 (t, J=7.1 Hz,3H).

Example 165a, b, c, d4-Cyano-N-((R)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide4-Cyano-N-((S)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide4-Cyano-N-((R)-1-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide4-Cyano-N-((S)-1-(trans-4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide(Homochiral with Absolute and Relative Stereochemistry not Determined)

Example 1654-Cyano-N-(1-(4-(6-fluoroquinolin-4-yl)cyclohexyl)propyl)benzamide

To a solution of 4-cyanobenzoic acid (33.4 mg, 0.227 mmol) in DMF (2 mL)was added HATU (86 mg, 0.227 mmol). The reaction mixture was stirred atrt for 10 min, followed by addition of1-(4-(6-fluoroquinolin-4-yl)cyclohexyl)propan-1-amine (50 mg, 0.175mmol) (Intermediate 164J) in THF (0.5 mL) and N-methyl morpholine (0.10mL, 0.838 mmol). The resulting mixture was stirred at rt for 2 h. Thereaction mixture was concentrated in vacuo and the residue was dissolvedin MeOH, filtered, and purified via preparative HPLC to give a mixturecontaining the four isomers. The isomers were further separated bypreparative SFC (Method K) to give:

First eluting Example 165a (10.7 mg, 0.025 mmol, 14.6% yield). LC-MSAnal. Calc'd for C₂₆H₂₆FN₃O 415.21 found [M+H] 416.2, T_(r)=1.40 min(Method B). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.82 (d, J=4.4 Hz, 1H), 8.37(d, J=9.1 Hz, 1H), 8.08 (dd, J=9.0, 5.8 Hz, 1H), 8.02-7.87 (m, 5H),7.69-7.58 (m, 1H), 7.46 (d, J=4.4 Hz, 1H), 4.29 (d, J=8.2 Hz, 1H),3.53-3.42 (m, 1H), 1.96-1.56 (m, 10H), 1.50-1.31 (m, 1H), 0.88 (t, J=7.2Hz, 3H).

Second eluting Example 165b (5.7 mg, 0.014 mmol, 7.8% yield). LC-MSAnal. Calc'd for C₂₆H₂₆FN₃O 415.21 found [M+H] 416.0, T_(r)=1.51 min(Method I). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.78 (d, J=4.5 Hz, 1H), 8.34(d, J=9.0 Hz, 1H), 8.13-7.99 (m, 3H), 7.99-7.86 (m, 3H), 7.74-7.57 (m,1H), 7.43 (d, J=4.4 Hz, 1H), 3.81 (br. s., 1H), 3.27 (t, J=11.4 Hz, 1H),2.00-1.81 (m, 4H), 1.76-1.30 (m, 7H), 0.87 (t, J=7.2 Hz, 3H).

Third eluting Example 165c (5.5 mg, 0.013 mmol, 7.5% yield). LC-MS Anal.Calc'd for C₂₆H₂₆FN₃O 415.21 found [M+H] 416.0, T_(r)=1.51 min (MethodI). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.79 (d, J=4.5 Hz, 1H), 8.34 (d, J=8.9Hz, 1H), 8.13-8.00 (m, 3H), 7.99-7.86 (m, 3H), 7.72-7.55 (m, 1H), 7.43(d, J=4.5 Hz, 1H), 3.82 (d, J=9.0 Hz, 1H), 3.28 (t, J=11.7 Hz, 1H),1.99-1.80 (m, 4H), 1.75-1.29 (m, 7H), 0.87 (t, J=7.2 Hz, 3H).

Fourth eluting Example 165d (12 mg, 0.029 mmol, 16.4% yield). LC-MSAnal. Calc'd for C₂₆H₂₆FN₃O 415.21 found [M+H] 416.0, T_(r)=1.52 min(Method I). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.81 (d, J=4.4 Hz, 1H), 8.38(d, J=9.1 Hz, 1H), 8.08 (dd, J=9.1, 5.8 Hz, 1H), 8.01-7.88 (m, 5H),7.70-7.60 (m, 1H), 7.47 (d, J=4.4 Hz, 1H), 4.28 (d, J=8.4 Hz, 1H),3.49-3.28 (m, 1H), 1.96-1.56 (m, 10H), 1.48-1.31 (m, 1H), 0.87 (t, J=7.2Hz, 3H).

Examples 176 to 196

Examples 176 to 196 were prepared from Intermediate 40L following theprocedure for Example 164 using the corresponding acid.

Ex. Tr No. Name R (min)^(Method 1*) [M + H]⁺ 1764-cyano-N-((R)-1-(cis-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

1.31 402.2 178 5-(3-fluoro-4-methoxyphenyl)-N-((R)-1-(cis-4-(6-fluoroquinolin-4- yl)cyclohexyl)ethyl)picolinamide

1.83 502.3 194 (R)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)-N-methylethanamine

1.40 459.9 195 N-((R)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(1H-pyrrol-1- yl)benzamide

1.56 442.0 196 N-((R)-1-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(1H-imidazol- 1-yl)benzamide

0.95 443.3 *unless otherwise noted

Example 1974-chloro-N-(1-(4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)ethyl)benzamide,4-chloro-N-((R)-1-(cis-4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)ethyl)benzamide,4-chloro-N-((S)-1-(cis-4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)ethyl)benzamide,4-chloro-N-((R)-1-(trans-4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)ethyl)benzamide,4-chloro-N-((S)-1-(trans-4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)ethyl)benzamideAbsolute and Relative Stereochemistry Unknown, Arbitrarily Assigned

197A. ethyl 2-(1,4-dioxaspiro[4.5]decan-8-ylidene)propanoate

To a suspension of NaH (0.307 g, 7.68 mmol) in THF (8 mL) cooled at 0°C. was added ethyl 2-(diethoxyphosphoryl)propanoate (1.830 g, 7.68 mmol)slowly. After 30 min, 1,4-dioxaspiro[4.5]decan-8-one (1 g, 6.40 mmol)was added. The resulting mixture was stirred at 0° C. for 2 hours, thenwarmed up to room temperature for overnight. The mixture was quenchedwith water, THF was removed under reduced pressure. The residue wasdissolved in EtOAc, washed with water, brine, dried over Na₂SO₄ andconcentrated. The crude was purified by ISCO (EtOAc/Hex 0-30%).Fractions containing the product were concentrated to yield Intermediate197A (1.2 g, 78% yield) a light yellow oil. ¹H NMR (400 MHz,CHLOROFORM-d) δ 4.19 (q, J=7.1 Hz, 2H), 4.03-3.89 (m, 4H), 2.68-2.53 (m,2H), 2.46-2.28 (m, 2H), 1.89 (s, 3H), 1.78-1.66 (m, 4H), 1.30 (t, J=7.1Hz, 3H)

197B. ethyl 2-(1,4-dioxaspiro[4.5]decan-8-yl)propanoate

A suspension of Intermediate 143A (500 mg, 2.081 mmol) (1A) and 10%palladium on carbon (25 mg, 0.024 mmol) in EtOAc (5 mL) was hydrogenatedin a Parr shaker at 45 psi for 6 h. The catalyst was filtered, and thefiltrate was concentrated to yield Intermediate 197B (450 mg, 89% yield)as a light oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.12 (dtt, J=10.7, 7.1,3.6 Hz, 2H), 3.98-3.81 (m, 4H), 2.35-2.17 (m, 1H), 1.83-1.68 (m, 3H),1.66-1.45 (m, 4H), 1.43-1.28 (m, 2H), 1.27-1.22 (m, 3H), 1.14-1.07 (m,3H)

197C. ethyl 2-(4-oxocyclohexyl)propanoate

To a solution of ethyl 2-(1,4-dioxaspiro[4.5]decan-8-yl)propanoate (450mg, 1.857 mmol) (1B) in THF (5 mL) was added 1M hydrogen chloride(aqueous) (0.929 mL, 3.71 mmol). The mixture was heated to 50° C. for 6h. The reaction mixture was concentrated. The residue was dissolved inEtOAc, washed with water (2×), brine, dried over Na₂SO₄ andconcentrated. The crude was purified with ISCO (EtOAc/Hex 0-30%).Fractions containing product were concentrated to yield Intermediate197C (290 mg, 79% yield) as a clear oil. ¹H NMR (400 MHz, CHLOROFORM-d)δ 4.22-4.06 (m, 2H), 2.46-2.30 (m, 5H), 2.13-1.91 (m, 3H), 1.56-1.42 (m,2H), 1.31-1.24 (m, 3H), 1.18 (d, J=7.1 Hz, 3H)

197D. ethyl2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)propanoate

Intermediate 143C (200 mg, 1.01 mmol) (1C) and2,6-di-tert-butyl-4-methylpyridine (238 mg, 1.16 mmol) were dissolved indry DCM (10 ml). To the reaction mixture trifluoromethanesulfonicanhydride (0.186 mL, 1.11 mmol) was added dropwise and stirred for 2 h.The suspension was filtered and the filtrate was diluted with DCM,washed with 1N HCl (2×), satd. sodium bicarb solution, water, brine anddried over Na₂SO₄ and concentrated to yield Intermediate 197D (320 mg,96% yield) as a brown oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.73 (t,J=6.1 Hz, 1H), 4.28-4.05 (m, 2H), 2.52-2.17 (m, 4H), 2.08-1.79 (m, 3H),1.49 (dt, J=11.1, 6.6 Hz, 1H), 1.31-1.20 (m, 3H), 1.19-1.04 (m, 3H)

197E. ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)propanoate

To a solution of Intermediate 143D (300 mg, 0.908 mmol) (1D) in DMSO (5mL) was added4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (230 mg,0.908 mmol) and potassium acetate (267 mg, 2.72 mmol). After the mixturewas degassed with N₂ for 10 min, PdCl₂(dppf) (19.9 mg, 0.027 mmol) wasadded. The mixture was heated to 80° C. for overnight. The mixture waspartitioned between EtOAc and water. The organic phase was concentratedand purified by ISCO. Fractions containing product were concentrated toyield Intermediate 197E (168 mg, 60% yield) as a brown oil. ¹H NMR (400MHz, CHLOROFORM-d) δ 6.66-6.40 (m, 1H), 4.31-4.00 (m, 2H), 2.34-2.26 (m,1H), 2.25-2.19 (m, 1H), 2.19-2.04 (m, 2H), 1.95-1.75 (m, 3H), 1.73-1.60(m, 1H), 1.29-1.24 (m, 15H), 1.13 (dd, J=11.6, 7.0 Hz, 3H)

197F. Ethyl2-(4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohex-3-en-1-yl)propanoate

A mixture of 7-chloropyrazolo[1,5-a]pyrimidine (0.193 g, 1.260 mmol),Intermediate 143E (0.400 g, 1.298 mmol), Na₂CO₃ (0.534 g, 5.04 mmol),and Pd(Ph₃P)₄ (0.073 g, 0.063 mmol) in dioxane (11.67 ml) and water(3.89 ml) was heated at 100° C. overnight. The reaction was quenchedwith water and diluted with EtOAc. Layers were separated. The aqueousphase was extracted with EtOAc (3×). The organics were combined, driedover Na₂SO₄, filtered, and concentrated to afford a brown residue.Purification of the crude material by silica gel chromatography using anISCO machine (40 g column, 40 mL/min, 0-70% EtOAc in hexanes over 16min, t_(r)=10.5 min) gave 197F (0.224 g, 0.748 mmol, 59.4% yield) as ayellow residue. ESI MS (M+H)⁺=300.2. HPLC Peak t_(r)=0.95 minutes. HPLCconditions: method A.

197G. Ethyl 2-(4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)propanoate

To a solution of 143F (0.224 g, 0.748 mmol) in MeOH (3.74 ml) was addedammonium formate (0.236 g, 3.74 mmol) followed by Pd/C (0.021 g, 0.202mmol). The reaction was heated at 70° C. for 1 h. The reaction wasfiltered through CELITE® and the filter cake washed with CH₂Cl₂. Thefiltrate was concentrated. The crude material was taken up in EtOAc andwashed with a sat. aq. solution of NaHCO₃ (1×). The organic phase wasdried over Na₂SO₄, filtered, and concentrated to afford 197G (220 mg,98%) as a yellow residue. ESI MS (M+H)⁺=302.2. HPLC Peak t_(r)=0.94minutes. HPLC conditions: method A.

197H. 2-(4-(Pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)propanoic acid

To a solution of 197G (0.1112 g, 0.369 mmol) in THF (1.318 ml) and MeOH(0.527 ml) was added lithium hydroxide (3.69 ml, 3.69 mmol). Thereaction was heated at 70° C. for 2.5 h, then allowed to cool to rt. Thereaction was adjusted to pH 7 with 1N HCl, then diluted with EtOAc.Layers were separated. The aqueous phase was extracted with EtOAc (5×).The organic phases were combined, dried over Na₂SO₄, filtered, andconcentrated to afford 197H (82.7 mg, 82%) as a yellow residue. ESI MS(M+H)⁺=274.1. HPLC Peak t_(r)=0.73 minutes. HPLC conditions: method A.

197I. 1-(4-(Pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)ethanamine

197H (0.0823 g, 0.301 mmol) was taken up in toluene (1.004 ml) in areaction vial and diphenyl phosphorazidate (0.071 ml, 0.331 mmol) andtriethylamine (0.050 ml, 0.361 mmol) were added. The vial was sealed andheated to 80° C. After about 2 h, the reaction was cooled to rt. Thecrude residue taken up in 1 mL THF and 1 mL of water and lithiumhydroxide (0.072 g, 3.01 mmol) were added. The reaction was stirred atrt. The reaction was acidified to pH=1 with 1N HCl and extracted withEtOAc to remove DPPA related impurities. The organic layer wasdiscarded. The aqueous layer was then basified to pH=12 with 1N NaOH andextracted with EtOAc (3×). The combined organic phases were dried withsodium sulfate, filtered, and concentrated in vacuo to give 197I (46.5mg, 0.190 mmol, 63.2% yield) as an orange residue. ESI MS (M+H)⁺=245.2.HPLC Peak t_(r)=0.52 minutes. HPLC conditions: method A.

Example 197a (+/−)-Cis- andtrans-4-chloro-N-(1-(4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)ethyl)benzamide

To a solution of 197I (46.5 mg, 0.190 mmol) in THF (1359 μl) at rt wasadded 4-chlorobenzoic acid (89 mg, 0.571 mmol), followed by1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (109 mg,0.571 mmol), 4-hydroxybenzotriazole (77 mg, 0.571 mmol) and Hunig's Base(133 μl, 0.761 mmol). The reaction was stirred at rt for 16 h. Thereaction was concentrated, then purified via preparative LC/MS with thefollowing conditions: Column: XBridge C18, 19×200 mm, 5-μm particles;Mobile Phase A: 5:95 acetonitrile: water with 10-mM ammonium acetate;Mobile Phase B: 95:5 acetonitrile: water with 10-mM ammonium acetate;Gradient: 20-70% B over 20 minutes, then a 5-minute hold at 100% B;Flow: 20 mL/min. Fractions containing the desired product were combinedand dried via centrifugal evaporation to afford the title compound as amixture of 4 isomers (22.5 mg, 30%). ESI MS (M+H)⁺=383.0. HPLC Peakt_(r)=1.714 minutes. Purity=98%. HPLC conditions: method B.

Approximately 20.6 mg of Example 197a was resolved by the followingmethod. The isomeric mixture was purified via preparative SFC with thefollowing conditions: Column: Chiral AD, 25×3 cm ID, 5-μm particles;Mobile Phase A: 70/30 CO₂/MeOH; Detector Wavelength: 220 nm; Flow: 85mL/min. The fractions (“Peak-1” t_(r)=7.485, “Peak-2” t_(r)=9.868,“Peak-3” t_(r)=11.635, “Peak-4” t_(r)=16.651; analytical conditions:Column: Chiral AD, 250×4.6 mm ID, 5-μm particles; Mobile Phase A: 70/30CO₂/MeOH; Flow: 2.0 mL/min) were collected in MeOH. The stereoisomericpurity of each fraction was estimated to be greater than 99% based onthe prep-SFC chromatograms. Each diasteromer or enantiomer was furtherpurified via preparative LC/MS:

Example 197b, first eluting isomer: The crude material was purified viapreparative LC/MS with the following conditions: Column: XBridge C18,19×200 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with10-mM ammonium acetate; Gradient: 20-70% B over 20 minutes, then a5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing thedesired product were combined and dried via centrifugal evaporation toafford Isomer 1 (5.0 mg, 6.6%). ESI MS (M+H)+=383.3. HPLC Peakt_(r)=1.764 minutes. Purity=96%. HPLC conditions: B. Absolutestereochemistry not determined.

Example 197c, second eluting isomer: The crude material was purified viapreparative LC/MS with the following conditions: Column: XBridge C18,19×200 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with10-mM ammonium acetate; Gradient: 20-70% B over 20 minutes, then a5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing thedesired product were combined and dried via centrifugal evaporation. Thematerial was further purified via preparative LC/MS with the followingconditions: Column: XBridge C18, 19×200 mm, 5-μm particles; Mobile PhaseA: 5:95 acetonitrile: water with 10-mM ammonium acetate; Mobile Phase B:95:5 acetonitrile: water with 10-mM ammonium acetate; Gradient: 35-65% Bover 25 minutes, then a 2-minute hold at 65% B; Flow: 20 mL/min.Fractions containing the desired product were combined and dried viacentrifugal evaporation to afford Isomer 2 (5.2 mg, 7.0%). ESI MS(M+H)+=383.1. HPLC Peak t_(r)=1.726 minutes. Purity=98%. HPLCconditions: B. Absolute stereochemistry not determined.

Example 197d, third eluting isomer: The crude material was purified viapreparative LC/MS with the following conditions: Column: XBridge C18,19×200 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with10-mM ammonium acetate; Gradient: 20-70% B over 20 minutes, then a5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing thedesired product were combined and dried via centrifugal evaporation toafford Isomer 3 (4.7 mg, 6.3%). ESI MS (M+H)+=383.2. HPLC Peakt_(r)=1.848 minutes. Purity=97%. HPLC conditions: B. Absolutestereochemistry not determined.

Example 197e, fourth eluting isomer: The crude material was purified viapreparative LC/MS with the following conditions: Column: XBridge C18,19×200 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with10-mM ammonium acetate; Gradient: 20-70% B over 20 minutes, then a5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing thedesired product were combined and dried via centrifugal evaporation toafford Isomer 4 (4.5 mg, 5.9%). ESI MS (M+H)+=383.2. HPLC Peakt_(r)=1.806 minutes. Purity=96%. HPLC conditions: B. Absolutestereochemistry not determined.

Example 1984-chloro-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)but-3-en-1-yl)benzamide

198A.(R)-3-((R)-2-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)pent-4-enoyl)-4-phenyloxazolidin-2-one

To a solution of Preparation 40I (50 mg, 0.116 mmol) in THF (2 mL) at−40° C. was added NaHMDS (1M in THF) (0.139 mL, 0.139 mmol) drop wise.The mixture was stirred at −40° C. to −30° C. for 15 min. Then3-bromoprop-1-ene (28.0 mg, 0.231 mmol) in THF (0.5 mL) was added dropwise. The reaction was stirred at −20° C. for 16 h. The reaction wasquenched at −20° C. by pouring it into saturated NH4Cl solution. Theaqueous was extracted with EtOAc. The organic was washed with brine,dried over MgSO₄, filtered and concentrated to give a crude material.This crude material was added MeOH and filtered to remove the solid. Thefiltrate was purified with prep HPLC (Phen Luna 5u 30×100 mm), 40 mL/minflow rate with gradient of 20% B-100% B over 10 minutes Hold at 100% Bfor 5 min. (A: 0.1% TFA in water/MeOH (90:10), B: 0.1% TFA in water/MeOH(10:90) monitoring at 254 nm. Combined fractions (tr=9.428 min)containing the product. After concentration, (R)-3-((R)-2-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)pent-4-enoyl)-4-phenyloxazolidin-2-one(25 mg, 0.052 mmol, 44.8% yield) was obtained as white solid. ¹H NMR(400 MHz, CHLOROFORM-d) δ 9.12 (d, J=5.5 Hz, 1H), 8.64 (dd, 5.0 Hz, 1H),8.01-7.89 (m, 2H), 7.89-7.75 (m, 1H), 7.47-7.31 (m, 5H), 5.62-5.45 (m,2H), 4.84-4.76 (m, 1H), 4.76-4.68 (m, 1H), 4.68-4.52 (m, 1H), 4.36 (dd,J=9.0, 3.9 Hz, 1H), 3.55-3.33 (m, 1H), 2.49-2.35 (m, 1H), 2.33-2.21 (m,2H), 2.12-1.97 (m, 2H), 1.93-1.65 (m, 6H) LC-MS: M+H=473.3 (tr=0.90 min)(Method A)

198B: (R)-2-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)pent-4-enoicacid

To a solution of Preparation 198A (250 mg, 0.529 mmol) in THF (2 mL) at0° C. was added 2.0 M LiOH in H₂O (0.476 mL, 0.952 mmol), followed by30% H₂O₂ (0.360 mL, 3.17 mmol). The reaction was stirred at 0° C. for 10min. Then it was warmed up to RT and stirred at RT for 16 h. Thereaction was carefully quenched at 0° C. by addition of saturatedNa2SO3. The pH was adjusted to 5˜6 with 1N HCl and the mixture wasextracted with EtOAc. The combined organics were dried over MgSO4,filtered and concentrated. The crude material was purified with prepHPLC (9 injections) (Phen Luna 5u 30×100 mm), 40 mL/min flow rate withgradient of 20% B-100% B over 10 minutes Hold at 100% B for 5 min. (A:0.1% TFA in water/MeOH (90:10), B: 0.1% TFA in water/MeOH (10:90)monitoring at 254 nm. 1B (78 mg, 0.236 mmol, 44.6% yield) was obtainedas white solid. ¹H NMR (400 MHz, CHLOROFORM-d) δ 9.22 (br. s., 1H), 8.63(dd, J=9.0, 5.0 Hz, 1H), 7.98-7.75 (m, 4H), 5.85 (dd, J=16.9, 9.7 Hz,1H), 5.25-5.03 (m, 2H), 3.50 (br. s., 1H), 2.89-2.75 (m, 1H), 2.54-2.32(m, 2H), 2.16 (d, J=10.1 Hz, 1H), 2.06 (d, J=13.2 Hz, 1H), 2.01-1.71 (m,6H) LC-MS: M+H=328 (tr=0.69 min) (Method A)

198C:(R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)but-3-en-1-amine

Preparation 198B (55 mg, 0.168 mmol) taken up in toluene (1 mL) anddiphenylphosphoryl azide (0.040 mL, 0.185 mmol) and triethylamine (0.028mL, 0.202 mmol) was added. Vial sealed and heated to 70 C. After about 3h, diphenylphosphoryl azide (0.040 mL, 0.185 mmol) and triethylamine(0.028 mL, 0.202 mmol) were added. The reaction was heated for another 3h. The reaction was cooled to rt and concentrated under reducedpressure. Crude residue taken up in THF (0.2 mL) and 2M LiOH (0.840 mL,1.680 mmol). Reaction stirred at rt for 16 h. LCMS shows isocyanateconsumed. New peak with M+1 of desired at rt=0.56 min. Reactionacidified with 1N HCl (white precipitate forms) to pH1 and extractedEtOAc to remove DPPA related impurities. The material was purified withprep HPLC (Phen Luna 5u 30×100 mm), 40 mL/min flow rate with gradient of0% B-100% B over 10 minutes Hold at 100% B for 5 min. (A: 0.1% TFA inwater/MeOH (90:10), B: 0.1% TFA in water/MeOH (10:90) monitoring at 254nm. Preparation 198C was obtained (30 mg, 0.040 mmol, 23.77% yield) wasobtained. LC-MS: M+H=299.2 (TR=0.56 min) (Method A).

Example 198 4-chloro-N-((R)-1-41 s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)but-3-en-1-yl)benzamide

To a solution of Preparation 198C (15 mg, 0.029 mmol) in THF (0.5 mL) atRT was added Hunig's Base (0.015 mL, 0.086 mmol), followed by4-chlorobenzoyl chloride (9.98 mg, 0.057 mmol). The reaction as stirredat RT for 2 h. The crude material was purified via preparative LC/MSwith the following conditions: Column: XBridge C18, 19×200 mm, 5-μmparticles; Mobile Phase A: 5:95 acetonitrile: water with 10-mM ammoniumacetate; Mobile Phase B: 95:5 acetonitrile: water with 10-mM ammoniumacetate; Gradient: 50-100% B over 20 minutes, then a 10-minute hold at100% B; Flow: 20 mL/min. Fractions containing the desired product werecombined and dried via centrifugal evaporation. The yield of the product1 was 3.8 mg (8.70 umol, 30.5%).

¹H NMR (500 MHz, DMSO-d₆) δ 8.82 (d, J=4.4 Hz, 1H), 8.27 (d, J=9.0 Hz,1H), 8.09 (dd, J=9.0, 5.9 Hz, 1H), 7.95 (d, J=10.9 Hz, 1H), 7.84 (d,J=8.3 Hz, 2H), 7.66 (t, J=7.5 Hz, 1H), 7.57-7.43 (m, 3H), 5.90-5.73 (m,1H), 5.07 (d, J=17.2 Hz, 1H), 4.98 (d, J=10.1 Hz, 1H), 4.42 (d, J=8.9Hz, 1H), 3.39 (br. s., 1H), 2.26-2.13 (m, 1H), 1.94-1.71 (m, 7H), 1.68(br. s., 1H), 1.62 (d, J=11.1 Hz, 1H) LC-MS: M+H=437.3 tr=2.23 min(Method B)

Example 199 N-((R)-1-((Is,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)but-3-en-1-yl)-[1,1′-biphenyl]-4-carboxamide

Example 199 was obtained following the procedures in Example 198 using198C and [1,1′-biphenyl]-4-carbonyl chloride. ¹H NMR (500 MHz, DMSO-d₆)δ 8.83 (d, J=4.5 Hz, 1H), 8.25 (d, J=9.2 Hz, 1H), 8.09 (dd, J=9.1, 5.8Hz, 1H), 8.01-7.84 (m, 3H), 7.80-7.61 (m, 5H), 7.54-7.45 (m, 3H),7.45-7.27 (m, 1H), 5.90-5.79 (m, 1H), 5.10 (d, J=17.4 Hz, 1H), 5.00 (d,J=9.8 Hz, 1H), 4.46 (d, J=7.9 Hz, 1H), 3.40 (br. s., 1H), 2.31-2.16 (m,1H), 2.01-1.78 (m, 6H), 1.75 (br. s., 2H), 1.69 (br. s., 1H), 1.63 (d,J=12.2 Hz, 1H) LC-MS: M+H=470.3 tr=2.41 min (Method B)

Example 200 and Example 201 (Chiral)N-1-((1s,4S)-4-(quinolin-3-yl)cyclohexyl)propyl)-[1,1′-biphenyl]-4-carboxamide

200A: ethyl 2-(4-(quinolin-3-yl)cyclohex-3-en-1-yl)acetate

Ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)acetate(5.26 g, 17.88 mmol) was taken up in Dioxane (40 mL) and Water (10.00mL). 3-bromoquinoline (3.1 g, 14.90 mmol) was added followed bypotassium carbonate (6.18 g, 44.7 mmol). Mixture was bubble with N2 for5 minutes before addition of tetrakis(triphenylphosphine)palladium(0)(0.344 g, 0.298 mmol). After addition, reaction was evacuated andbackfilled with N₂ three times and then sealed and heated to 100° C. for16 h. The Reaction was diluted with EtOAc and water. Organic wasseparated and washed with brine, dried over MgSO₄, filtered andconcentrated in vacuo and purified directly via ISCO (120 g column, 85mL/min, 0-30% EtOAc in hexanes) to give Preparation 200A (4.47 g, 14.38mmol, 96% yield). 1H NMR (400 MHz, CHLOROFORM-d) δ 9.05 (d, J=2.3 Hz,1H), 8.09 (d, J=8.4 Hz, 1H), 8.02 (d, J=2.2 Hz, 1H), 7.86-7.76 (m, 1H),7.67 (ddd, J=8.4, 6.9, 1.5 Hz, 1H), 7.58-7.45 (m, 1H), 6.38-6.18 (m,1H), 4.20 (q, J=7.1 Hz, 2H), 2.67-2.56 (m, 2H), 2.55-2.43 (m, 1H),2.42-2.35 (m, 2H), 2.30-2.18 (m, 1H), 2.12-1.92 (m, 2H), 1.57 (ddt,J=12.8, 10.8, 7.9 Hz, 1H), 1.36-1.27 (m, 3H) LC-MS: M+H=296.2 tr=0.74min (Method A)

200B: ethyl 2-(4-(quinolin-3-yl)cyclohexyl)acetate

Preparation 200A (3.5 g, 11.85 mmol) was dissolved in MeOH (70 mL) andammonium formate (3.74 g, 59.2 mmol) was added. The vessel was equippedwith a reflux condenser and vacated and flushed with N₂ 3 times. Then,10% Pd/C (1.256 g, 1.185 mmol) was added and the reaction was heated at70° C. LCMS after 1 hour shows reduction complete. Reaction cooled,solids were filtered off and the filtrate was concentrated to give crudematerial. This crude material was purified with ISCO 120 g, 85 mL/min.0-50% EtOAc/Hexane. Preparation 200B (0.71 g, 2.308 mmol, 19.48% yield)was eluted with 10% EtOAc/Hexane. LC-MS: M+H=302.2 tr=0.81 min (MethodA)

200C: ethyl 2-(4-(quinolin-3-yl)cyclohexyl)butanoate

To a solution of Preparation 200B (920 mg, 3.09 mmol) in THF (10 mL) at0° C. was added 1M NaHMDS in THF (7.73 mL, 7.73 mmol) drop wise. Themixture was stirred at 0° C. for 30 min. Then iodoethane (0.3 mL, 3.75mmol) was added drop wise. The resulting mixture was stirred at 0° C.for 45 min. [The color of the solution does not change much]. Iodoethane(0.4 mL, 5.00 mmol) was added drop wise and the reaction was stirred at0° C. [The color of the solution turned to a little darker]. After 1 h,iodoethane (0.15 mL, 1.875 mmol) was added and the reaction was stirredat RT for 2 h. LC-MS shows desired product formed but still there isstarting material left. The reaction was poured into a saturated NH₄Clsolution. EtOAc was added and organic was separated and washed withbrine, dried over MgSO₄, filtered and concentrated to give a crudematerial. This crude material was purified with ISCO 80 g column, 60mL/min. 0-30% EtOAc/Hexane in 40 min. The desired product was elutedwith 25% EtOAc/Hexane. Combined fractions 5-11. After concentration,Preparation 200C (386 mg, 1.174 mmol, 38.0% yield) was obtained as clearliquid. LC-MS; M+H=326.2 (TR=0.81, 0.82 min) (Method A)

200D: 2-(4-(quinolin-3-yl)cyclohexyl)butanoic acid

To a solution of Preparation 200C (385 mg, 1.183 mmol) in THF (1 mL) andMeOH (5 mL) at rt was added 2M LiOH (5.91 mL, 11.83 mmol) and 1M NaOH(2.366 mL, 2.366 mmol). The reaction was stirred at 60° C. for 48 h.LC-MS still shows a little bit of starting material and methylester.Cooled to RT. The mixture was adjusted to pH 5 with concentrated HCl.Extracted the aqueous layer with EtOAc. The organic was separated andwashed with brine, dried over MgSO₄, filtered and concentrated to givePreparation 200D (400 mg, 1.076 mmol, 91% yield) as white solid. LC-MS:M+H=298.2 (tr=0.67 min) (Method A)

200E: 1-(4-(quinolin-3-yl)cyclohexyl)propan-1-amine

Preparation 200D (200 mg, 0.673 mmol) taken up in toluene (2 mL) anddiphenylphosphoryl azide (0.290 mL, 1.345 mmol) and triethylamine (0.187mL, 1.345 mmol) added. Vial sealed and heated to 70 C for 16 h. Thereaction was cooled to RT and concentrated under reduced pressure. Cruderesidue taken up in THF (2 mL) and 2M LiOH (2.354 mL, 4.71 mmol).Reaction stirred at RT for 3 days. LCMS shows isocyanate consumed. Newpeak with M+1 of desired at RT=0.50 min. Reaction acidified with 1N HClto pH1 and extracted EtOAc to remove DPPA related impurities. Then theaqueous layer was adjusted to pH 10 with 2M LiOH. Extracted with EtOAc(3×). Combined organics were washed with brine, dried over MgSO₄,filtered and concentrated to give Preparation 200E (110 mg, 0.398 mmol,59.1% yield) as clear liquid. LC-MS: M+H=269.2 (tr=0.50 min) (Method A)

Example 200a and Example 200bN-(1-((1r,4r)-4-(quinolin-3-yl)cyclohexyl)propyl)-[1,1′-biphenyl]-4-carboxamideandN-(1-((1r,4r)-4-(quinolin-3-yl)cyclohexyl)propyl)-[1,1′-biphenyl]-4-carboxamide(Relative and Absolute Stereochemistry No Confirmed, ArbitrarilyAssigned)

To a solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (102 mg, 0.533 mmol) in DMF (4 mL) at RT was added[1,1′-biphenyl]-4-carboxylic acid (162 mg, 0.820 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (102 mg,0.533 mmol),1-hydroxybenzotriazole (82 mg, 0.533 mmol) and triethylamine(0.171 mL, 1.230 mmol). The reaction was stirred at RT for 16 h. Thereaction was diluted with EtOAc and water. Organic was separated andwashed with brine, dried over MgSO₄, filtered and concentrated todryness. The crude material was purified with ISCO 40 g column, 40mL/min, 0-70% EtOAc/Hexane in 40 min. 3F (90 mg, 0.197 mmol, 48%) waseluted with 60% EtOAc/Hexane.

Example 200a ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.86 (d, J=2.2 Hz, 1H),8.15-8.01 (m, 2H), 7.89-7.77 (m, 3H), 7.74-7.58 (m, 5H), 7.58-7.47 (m,3H), 7.47-7.34 (m, 1H), 5.81 (d, J=9.8 Hz, 1H), 4.56-4.27 (m, 1H), 2.96(br. s., 1H), 2.26-2.09 (m, 1H), 2.01-1.78 (m, 8H), 1.77-1.64 (m, 1H),1.52-1.36 (m, 1H), 1.08-0.96 (m, 3H) LC-MS: M+H=449.3 (tr=0.88 min)(Method A)

Example 200b (65 mg, 0.142 mmol, 35%) was eluted with 70% EtOAc/Hexane.¹H NMR (400 MHz, CHLOROFORM-d) δ 8.83 (d, J=2.3 Hz, 1H), 8.09 (d, J=8.4Hz, 1H), 7.96-7.85 (m, 3H), 7.79 (d, J=8.3 Hz, 1H), 7.73-7.61 (m, 5H),7.58-7.47 (m, 3H), 7.45-7.31 (m, 1H), 5.92 (d, J=9.5 Hz, 1H), 4.21-4.01(m, 1H), 2.82-2.55 (m, 1H), 2.18-1.95 (m, 4H), 1.83 (ddd, J=14.0, 7.4,4.5 Hz, 11-1), 1.75-1.47 (m, 4H), 1.47-1.32 (m, 2H), 1.05 (t, J=7.4 Hz,3H) LC-MS: M+H=449.3 (tr=0.88 min) (Method A)

Examples 200c and 200dN-((R)-1-((1s,4S)-4-(quinolin-3-yl)cyclohexyl)propyl)-[1,1′-biphenyl]-4-carboxamideandN-((S)-1-((1s,4R)-4-(quinolin-3-yl)cyclohexyl)propyl)-[1,1′-biphenyl]-4-carboxamide(Absolute and Relative Stereochemistry not Confirmed, ArbitrarilyAssigned)

The racemate Example 200a was purified via preparative SFC with thefollowing conditions: Column: Chiral AD-H 25×3 cm ID, 5-μm particles;Mobile Phase A: 50/50 CO2/MeOH; Detector Wavelength: 220 nm; Flow: 85mL/min. The fractions (“Peak-1” tr=10.78 min (Example 200c) and “Peak-2”tr=23.917 min (Example 200d);

¹H NMR (400 MHz, CHLOROFORM-d) δ 8.86 (d, J=2.2 Hz, 1H), 8.15-8.01 (m,2H), 7.89-7.77 (m, 3H), 7.74-7.58 (m, 5H), 7.58-7.47 (m, 3H), 7.47-7.34(m, 1H), 5.81 (d, J=9.8 Hz, 1H), 4.56-4.27 (m, 1H), 2.96 (br. s., 1H),2.26-2.09 (m, 1H), 2.01-1.78 (m, 8H), 1.77-1.64 (m, 1H), 1.52-1.36 (m,1H), 1.08-0.96 (m, 3H) LC-MS: M+H=449.3 (tr=0.88 min) (Method A)

Examples 201 and 202N-((R)-1-((1r,4R)-4-(quinolin-3-yl)cyclohexyl)propyl)-[1,1′-biphenyl]-4-carboxamideandN-((S)-1-((1r,4S)-4-(quinolin-3-yl)cyclohexyl)propyl)-[1,1′-biphenyl]-4-carboxamide(Absolute and Relative Stereochemistry not Confirmed, ArbitrarilyAssigned)

The racemate Example 200b was purified via preparative SFC with thefollowing conditions: Column: Chiral IC-H 25×3 cm ID, 5-μm particles;Mobile Phase A: 50/50 CO2/MeOH; Detector Wavelength: 220 nm; Flow: 85mL/min. The fractions (“Peak-1” tr=10.811 min (Example 201) and “Peak-2”tr=10.842 min (Example 202);

¹H NMR (400 MHz, CHLOROFORM-d) δ 8.83 (d, J=2.3 Hz, 1H), 8.09 (d, J=8.4Hz, 1H), 7.96-7.85 (m, 3H), 7.79 (d, J=8.3 Hz, 1H), 7.73-7.61 (m, 5H),7.58-7.47 (m, 3H), 7.45-7.31 (m, 1H), 5.92 (d, J=9.5 Hz, 1H), 4.21-4.01(m, 1H), 2.82-2.55 (m, 1H), 2.18-1.95 (m, 4H), 1.83 (ddd, J=14.0, 7.4,4.5 Hz, 1H), 1.75-1.47 (m, 4H), 1.47-1.32 (m, 2H), 1.05 (t, J=7.4 Hz,3H) LC-MS: M+H=449.2 (tr=0.86 min) (Method A)

Example 2034-chloro-N-((R)-1-(cis-4-(quinolin-3-yl)cyclohexyl)propyl)benzamide4-chloro-N-((S)-1-(cis-4-(quinolin-3-yl)cyclohexyl)propyl)benzamide4-chloro-N-((R)-1-(trans-4-(quinolin-3-yl)cyclohexyl)propyl)benzamide4-chloro-N-((S)-1-(trans-4-(quinolin-3-yl)cyclohexyl)propyl)benzamideAbsolute and Relative Stereochemistry Unknown

Examples 203a-d were obtained following the procedures in Examples 200and 201 using 200E and 4-chlorobenzoyl chloride. The racemate waspurified via preparative SFC with the following conditions: Column:Chiral IC-H 25×3 cm ID, 5-μm particles; Mobile Phase A: 65/35 CO2/MeOH;Detector Wavelength: 220 nm; Flow: 85 mL/min.

The fractions (“Peak-1” tr=5.98 min (Example 203a) and “Peak-2” tr=6.29min (Example 203b); (“Peak-3” tr=8.0 min (Example 203c) and “Peak-4”tr=9.0 min (Example 203d);

Examples 203a and 203b ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.86 (d, J=2.2Hz, 1H), 8.16-8.00 (m, 2H), 7.80 (d, J=8.2 Hz, 1H), 7.73-7.63 (m, 3H),7.59-7.46 (m, 1H), 7.45-7.35 (m, 2H), 5.71 (d, J=10.0 Hz, 1H), 4.49-4.23(m, 1H), 3.06-2.83 (m, 1H), 2.24-2.05 (m, 1H), 2.00-1.83 (m, 5H),1.83-1.73 (m, 3H), 1.73-1.63 (m, 1H), 1.51-1.35 (m, 1H), 1.00 (t, J=7.4Hz, 3H) LC-MS: M+H=407.2 (tr=0.81 min) (Method A)

Examples 203c and 203d ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.86 (d, J=2.2Hz, 1H), 8.16-8.00 (m, 2H), 7.80 (d, J=8.2 Hz, 1H), 7.73-7.63 (m, 3H),7.59-7.46 (m, 1H), 7.45-7.35 (m, 2H), 5.71 (d, J=10.0 Hz, 1H), 4.49-4.23(m, 1H), 3.06-2.83 (m, 1H), 2.24-2.05 (m, 1H), 2.00-1.83 (m, 5H),1.83-1.73 (m, 3H), 1.73-1.63 (m, 1H), 1.51-1.35 (m, 1H), 1.00 (t, J=7.4Hz, 3H) LC-MS: M+H=407.2 (tr=0.81 min) (Method A)

Example 207rac-4-chloro-N-(1-((trans)-4-((3-chloro-2-methylpyridin-4-yl)oxy)cyclohexyl)ethyl)benzamide

207A. rac-ethyl2-((trans)-4-((3-chloro-2-methylpyridin-4-yl)oxy)cyclohexyl)propanoate

A solution of ethyl rac-2-((trans)-4-hydroxycyclohexyl)propanoate (1.001g, 5 mmol) in THF (4 mL) was cooled to 0° C. and treated with potassiumhexamethyldisilazide (5.50 mL, 5.50 mmol) over 1 min. The reaction wasstirred 10 min. then treated with 3,4-dichloro-2-methylpyridine (0.851g, 5.25 mmol). The reaction was stirred 40 min. at 0° C. then quenchedwith aq. ammonium chloride. The phases were stirred together 1 h thenextracted with 1:1 EtOAc-hexane, and the organic extract was dried andstripped to afford an oil. Prep. HPLC afforded rac-ethyl2-((trans)-4-((3-chloro-2-methylpyridin-4-yl)oxy)cyclohexyl)propanoate(0.47 g, 29% yield) as a golden oil. MS (ES): m/z=326 [M+H]⁺. t_(R)=0.78min (Method A).

207B.rac-2-((trans)-4-((3-chloro-2-methylpyridin-4-yl)oxy)cyclohexyl)propanoicacid

A solution of Preparation 207A (0.42 g, 1.289 mmol) in THF (4 mL) wastreated with lithium hydroxide (0.154 g, 6.45 mmol) in water (4 mL).Methanol, ˜4 mL was added to give a single phase, and the reaction wasstirred for 1 h at 50° C. The reaction was then cooled and stirred atRT. Most of the solvent was removed under a stream of nitrogen, and thereaction was diluted to ˜6 ml with water. This cloudy suspension wasfiltered, and the filtrate solution pH was adjusted to ˜5.5 with aq.HOAc. The resulting precipitate was filtered, rinsed with water, andair-dried to affordrac-2-((trans)-4-((3-chloro-2-methylpyridin-4-yl)oxy)cyclohexyl)propanoicacid (0.16 g, 42% yield) as a white solid. MS (ES): m/z=298 [M+H]⁺.t_(R)=0.63 min (Method A).

207C.rac-1-((trans)-4-((3-chloro-2-methylpyridin-4-yl)oxy)cyclohexyl)ethanamine

A solution of Preparation 207B (0.26 g, 0.873 mmol) in toluene (4.37 ml)was treated with triethylamine (0.158 ml, 1.135 mmol) followed bydiphenylphosphinyl azide (0.244 g, 1.004 mmol). The solution was warmedto 70° C. (much bubbling). After 30 min., the solution was cooled andstripped. The residue was re-dissolved in THF (5 mL) and added to asolution of lithium hydroxide (0.836 g, 34.9 mmol) in 20 mL of water and8 mL of THF. This mixture was stirred at RT for 30 min. then it wasdiluted with ether and washed twice with 1M aq. HCl. The combinedaqueous phases were drained into sat. aq. sodium carbonate (finalpH-12), and this mixture was ext. with EtOAc then 3:1 chloroform-IPA.These two organic extracts were combined, dried, and stripped to affordrac-1-((trans)-4-((3-chloro-2-methylpyridin-4-yl)oxy)cyclohexyl)ethanamine(0.18 g, 77% yield) an oil. MS (ES): m/z=269 [M+H]⁺. t_(R)=0.47 min(Method A).

Example 207rac-4-chloro-N-(1-((trans)-4-((3-chloro-2-methylpyridin-4-yl)oxy)cyclohexyl)ethyl)benzamide

A solution of Preparation 207C (0.01 g, 0.037 mmol) and 4-chlorobenzoicacid (6.99 mg, 0.045 mmol) in DMF (0.25 mL) was treated withtriethylamine (0.016 mL, 0.112 mmol) followed by BOP (0.021 g, 0.048mmol). The reaction was stirred 2 h at RT then quenched with one drop ofwater and diluted with DMF to 2 mL. This solution was then purified byprep. HPLC. Concentration of the appropriate fractions afforded 0.0088 g(50%) of the title compound. MS (ES): m/z=407 [M+H]⁺. t_(R)=2.05 min(Method B).

Examples 208-210

Bop coupling (Scheme 9, below) of amine 207C (prepared in the preceedingexample) with the appropriate benzoic acids under the conditionsdescribed for the conversion of 207C to Example 207 affords compounds ofthe invention shown in Table 1 below. (All entries are racemic withtrans relative stereochemistry at the cyclohexyl.)

TABLE 1

Ex.# R (M + H)⁺ t_(R) (min., Method B) BMT# Example 208 F 391 1.93 BMT-267222 Example 209 OMe 403 1.83 BMT- 267223 Example 210 Me 387 1.96 BMT-267225

TABLE 2 Examples 211-225 were prepared following the procedures inExample 157 using the corresponding pyridyl halide (absolute andrelative stereochemistry unknown)

Example 211 4-chloro-N-(1-(4-(2- (trifluoromethyl)pyridin-4-yl)cyclohexyl)ethyl)benzamide

2.194^(B) 411.1 Diastereomer Mixture Example 212 4-chloro-N-(1-(4-(2-(trifluoromethyl)pyridin-4- yl)cyclohexyl)ethyl)benzamide

9.502^(AC) 411.3 Homochiral Example 213 4-chloro-N-(1-(4-(2-(trifluoromethyl)pyridin-4- yl)cyclohexyl)ethyl)benzamide

11.583^(AC) 411.3 Homochiral Example 214 4-chloro-N-(1-(4-(2-(trifluoromethyl)pyridin-4- yl)cyclohexyl)ethyl)benzamide

12.382^(AC) 411.3 Homochiral Example 215 4-chloro-N-(1-(4-(2-(trifluoromethyl)pyridin-4- yl)cyclohexyl)ethyl)benzamide

13.169^(AC) 411.3 Homochiral Example 216 4-chloro-N-(1-(4-(6-(trifluoromethyl)pyridin-3- yl)cyclohexyl)ethyl)benzamide

2.169^(B) 411.1 Diastereomer Mixture Example 217 4-chloro-N-(1-(4-(6-(trifluoromethyl)pyridin-3- yl)cyclohexyl)ethyl)benzamide

8.665^(AD) 411.1 Homochiral Example 218 4-chloro-N-(1-(4-(6-(trifluoromethyl)pyridin-3- yl)cyclohexyl)ethyl)benzamide

9.315^(AD) 411.1 Homochiral Example 219 4-chloro-N-(1-(4-(6-(trifluoromethyl)pyridin-3- yl)cyclohexyl)ethyl)benzamide

11.473^(AD) 411.1 Homochiral Example 220 4-chloro-N-(1-(4-(6-(trifluoromethyl)pyridin-3- yl)cyclohexyl)ethyl)benzamide

14.545^(AD) 411.1 Homochiral Example 221 N-(1-(4-(2-fluoro-3-methylpyridin-4- yl)cyclohexyl)ethyl)-[1,1′- biphenyl]-4-carboxamide

2.310^(B) 417.2 Diastereomer Mixture Example 222 N-(1-(4-(2-fluoro-3-methylpyridin-4- yl)cyclohexyl)ethyl)-[1,1′- biphenyl]-4-carboxamide

12.645^(AE) 417.2 Homochiral Example 223 N-(1-(4-(2-fluoro-3-methylpyridin-4- yl)cyclohexyl)ethyl)-[1,1′- biphenyl]-4-carboxamide

14.189^(AE) 417.2 Homochiral Example 224 N-(1-(4-(2-fluoro-3-methylpyridin-4- yl)cyclohexyl)ethyl)-[1,1′- biphenyl]-4-carboxamide

15.726^(AE) 417.2 Homochiral Example 225 N-(1-(4-(2-fluoro-3-methylpyridin-4- yl)cyclohexyl)ethyl)-[1,1′- biphenyl]-4-carboxamide

21.565^(AE) 417.2 Homochiral

Example 2264-Chloro-N-((1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)methyl)benzamide

226A. tert-Butyl4-((4-chlorobenzamido)methyl)-4-methylpiperidine-1-carboxylate

To a homogeneous mixture of tert-butyl4-(aminomethyl)-4-methylpiperidine-1-carboxylate (53.0 mg, 0.23 mmol) inanhydrous DCM (2 mL), under nitrogen atmosphere, was added DIPEA (0.17mL, 0.97 mmol) followed by 4-chlorobenzoyl chloride (0.05 mL, 0.390mmol). The resulting mixture was stirred at ambient temperature for 4hours, before being partitioned between DCM and water. The layers wereseparated and the aqueous layer was extracted twice more with DCM. Theseorganic extracts were combined with the original organic layer and wereconcentrated in vacuo to afford the title compound as an amber residue,which was used in the next step without purification. MS(ES): m/z=367[M+H]⁺. t_(R)=1.00 min (Method A).

226B. 4-Chloro-N-((4-methylpiperidin-4-yl)methyl)benzamide

To a homogeneous mixture of tert-butyl4-((4-chlorobenzamido)methyl)-4-methylpiperidine-1-carboxylate (226A,0.23 mmol) in anhydrous dioxane (3 mL), under nitrogen atmosphere, wasadded HCl (4N in dioxane, 0.5 mL, 2.0 mmol). The resulting mixture wasstirred at ambient temperature for 45 hours before being partitionedbetween water and EtOAc. The layers were separated and the aqueous layerwas extracted once more with EtOAc. The organic layers were combined andwashed with water, and this aqueous layer was added to the originalaqueous layer. The combined aqueous layer was lyophilized to afford theHCl salt of title compound as a brown residue which was used withoutfurther purification. MS (ES): m/z=267 [M+H]⁺. t_(R)=0.59 min (MethodA).

Example 2264-Chloro-N-((1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)methyl)benzamide

To a sealable flask charged with 4-chloro-6-fluoroquinoline (15.0 mg,0.08 mmol) was added a homogeneous mixture of the HCl salt of4-chloro-N-((4-methylpiperidin-4-yl)methyl)benzamide (226B, 23.4 mg,0.09 mmol) and DIPEA (0.07 mL, 0.40 mmol) in anhydrous NMP (1 mL). Thevial was sealed and the mixture was stirred at 120° C. After 65 hours,the reaction mixture was cooled to room temperature, diluted with DMF,passed through a syringe filter then purified via preparative HPLC/MS toafford the title compound (19.4 mg; 57% yield). MS(ES): m/z=412 [M+H]⁺.t_(R)=1.91 min (Method B). ¹H NMR (500 MHz, DMSO-d₆) δ 8.63-8.53 (m,2H), 8.00 (dd, J=9.1, 5.3 Hz, 1H), 7.85 (d, J=8.4 Hz, 2H), 7.81-7.71 (m,2H), 7.52 (d, J=8.3 Hz, 2H), 7.10 (d, J=6.3 Hz, 1H), 3.71-3.60 (m, 1H),3.55-3.43 (m, 1H), 3.31 (d, J=6.1 Hz, 2H), 2.95-2.85 (m, 1H), 2.56-2.54(m, 1H), 1.79-1.68 (m, 2H), 1.58-1.49 (m, 2H), 1.04 (s, 3H).

Example 2274-Chloro-N-((4-methyl-1-(2-(trifluoromethyl)pyridin-4-yl)piperidin-4-yl)methyl)benzamide

Example 227 (13.9 mg; 41% yield) was prepared following a procedureanalogous to that for the synthesis of Example 226 except that4-chloro-2-(trifluoromethyl)pyridine was used instead of4-chloro-6-fluoroquinoline, in the final step. MS(ES): m/z=412 [M+H]⁺.t_(R)=1.96 min (Method B). ¹H NMR (500 MHz, DMSO-d₆) δ 8.57-8.48 (m,1H), 8.19 (d, J=5.9 Hz, 1H), 7.78 (d, J=8.3 Hz, 2H), 7.49 (d, J=8.4 Hz,2H), 7.13 (s, 1H), 7.01-6.91 (m, 1H), 3.74-3.54 (m, 2H), 3.34-3.24 (m,2H), 3.21 (d, J=6.2 Hz, 2H), 1.55-1.43 (m, 2H), 1.39-1.30 (m, 2H), 0.96(s, 3H).

Example 228N-((4-Methyl-1-(2-(trifluoromethyl)pyridin-4-yl)piperidin-4-yl)methyl)-[1,1′-biphenyl]-4-carboxamide

Example 228 (15.6 mg; 44% yield) was prepared following a procedureanalogous to that for the synthesis of Example 227 except that[1,1′-biphenyl]-4-carbonyl chloride was used instead of 4-chlorobenzoylchloride, in the initial step. MS(ES): m/z=454 [M+H]⁺. t_(R)=2.12 min(Method B). ¹H NMR (500 MHz, DMSO-d₆) δ 8.49 (t, Hz, 1H), 8.21 (d, J=6.0Hz, 1H), 7.90 (d, J=8.2 Hz, 2H), 7.79-7.65 (m, 4H), 7.48 (t, J=7.5 Hz,2H), 7.44-7.36 (m, 1H), 7.16 (s, 1H), 7.04-6.94 (m, 1H), 3.69-3.53 (m,2H), 3.31 (t, J=9.8 Hz, 2H), 3.25 (d, J=6.1 Hz, 2H), 1.59-1.48 (m, 2H),1.41-1.32 (m, 2H), 0.99 (s, 3H).

Example 229(+/−)-4-chloro-N-(1-((1r,4r)-4-((2-(trifluoromethyl)pyridin-4-yl)oxy)cyclohexyl)ethyl)benzamide

Preparation 229A. ethyl2-((1r,4r)-4-((2-(trifluoromethyl)pyridin-4-yl)oxy)cyclohexyl)propanoate

To a solution of ethyl 2-((1r,4r)-4-hydroxycyclohexyl)propanoate (0.1294g, 0.646 mmol) in DMF (1.077 ml) was added NaH (0.043 g, 1.077 mmol).After 30 min, 4-bromo-2-(trifluoromethyl)pyridine (0.071 ml, 0.538 mmol)as added. The reaction was heated at 80° C. overnight. Reaction quenchedwith a sat. aq. soln of NH₄Cl and diluted with EtOAc. Layers wereseparated. The aqueous phase was extracted with EtOAc (2×). The combinedorganic phases were washed with water, dried over Na₂SO₄, filtered, andconcentrated to afford a brown residue. Purification of the crudematerial by silica gel chromatography using an ISCO machine (40 gcolumn, 40 mL/min, 0-30% EtOAc in hexanes over 14 min, t_(r)=9.5 min)gave the title compound (0.0646 g, 0.187 mmol, 34.7% yield) as acolorless residue. ESI MS (M+H)+=346.2. HPLC Peak tr=1.09 minutes. HPLCconditions: A.

Preparation 229B.2-((1r,4r)-4-((2-(trifluoromethyl)pyridin-4-yl)oxy)cyclohexyl)propanoicacid

To a solution of Preparation 229A (0.0437 g, 0.127 mmol) in THF (0.452ml) and MeOH (0.181 ml) was added lithium hydroxide (1.265 ml, 1.265mmol). The reaction was heated at 70° C. for 2 h, then allowed to coolto rt. The reaction was adjusted to pH 7 with 1N HCl, then diluted withEtOAc. Layers were separated. The aqueous phase was extracted with EtOAc(3×). The organic phases were combined, dried over Na₂SO₄, filtered, andconcentrated to afford the title compound as a colorless residue (18.2mg, 45% yield). ESI MS (M+H)+=318.1. HPLC Peak t_(r)=0.89 minutes. HPLCconditions: A.

Preparation 229C.1-((1r,4r)-4-((2-(trifluoromethyl)pyridin-4-yl)oxy)cyclohexyl)ethanamine

Preparation 229B (18.2 mg, 0.057 mmol) taken up in toluene (191 μl) anddiphenyl phosphorazidate (13.59 μl, 0.063 mmol) and triethylamine (9.59μl, 0.069 mmol) added. The vial was sealed and heated to 80° C. Afterabout 2 h, the reaction was cooled to rt. The reaction heated anaddition 2 h, then allowed to cool to rt. To this reaction was added 1mL THF and 1 mL of water and lithium hydroxide (13.74 mg, 0.574 mmol).The reaction stirred at rt overnight. The reaction was acidified to pH=1with 1N HCl (˜5.5 mL) and extracted with EtOAc to remove DPPA relatedimpurities. Then, the aqueous phase was basified to pH=12 with 1N NaOHand extracted with EtOAc (3×). The organic extracts were dried withsodium sulfate, filtered, and concentrated in vacuo to give the titlecompound (3.8 mg, 0.013 mmol, 22.98% yield) as a yellow residue.

Example 229(+/−)-4-chloro-N-(1-((1r,4r)-4-((2-(trifluoromethyl)pyridin-4-yl)oxy)cyclohexyl)ethyl)benzamide

To a solution of Preparation 229C (3.8 mg, 0.013 mmol) in THF (132 μl)at rt was added Hunig's base (6.91 μl, 0.040 mmol), followed by4-chlorobenzoyl chloride (3.38 μl, 0.026 mmol). The reaction was stirredat rt for 2 h. The crude material was purified via preparative LC/MSwith the following conditions: Column: XBridge C18, 19×150 mm, 5-μmparticles; Mobile Phase A: 5:95 acetonitrile: water with 10-mM ammoniumacetate; Mobile Phase B: 95:5 acetonitrile: water with 10-mM ammoniumacetate; Gradient: 25-100% B over 20 minutes, then a 5-minute hold at100% B; Flow: 20 mL/min. Fractions containing the desired product werecombined and dried via centrifugal evaporation to afford the titlecompound (2.5 mg, 44%). ESI MS (M+H)+=427.2. HPLC Peak t_(r)=2.101minutes. Purity=100%. HPLC conditions: B.

Example 230N-(1-((1s,4s)-4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)propyl)biphenyl-4-carboxamide

230A. ethyl2-(4-(6-(trifluoromethyl)quinolin-4-yl)cyclohex-3-enyl)acetate

To a solution of 4-chloro-6-(trifluoromethyl)quinoline (2.05 g, 8.85mmol), ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)acetate(3.12 g, 10.62 mmol) in 1,4-dioxane (35 mL) was added potassiumcarbonate (3.67 g, 26.6 mmol) and water (7 mL). The reaction mixture waspurged with nitrogen stream for 3 min, followed by addition of Pd(Ph₃P)₄(0.409 g, 0.354 mmol). The resulting mixture was heated at 100° C. undernitrogen stream for over night. The reaction mixture was cooled down anddiluted with ethyl acetate and saturated NaHCO₃ solution. The organiclayer was separated and washed with sat. NaHCO₃ solution, and dried overMgSO₄. The filtrate was concentrated in vacuo and the residue waspurified via silica gel flash column chromatography, eluting with 0-50%ethyl acetate in hexane to give Intermediate 230A (oil, 3.0 g, 8.26mmol, 93% yield). LC-MS Anal. Calc'd for C₂₀H₂₀F₃NO₂, 363.14. found[M+H] 364.5. T_(r)=0.97 min (Method A). ¹H NMR (400 MHz, CHLOROFORM-d)δ: 8.95 (d, J=4.5 Hz, 1H), 8.31 (s, 1H), 8.22 (d, J=8.8 Hz, 1H), 7.87(dd, J=8.8, 2.0 Hz, 1H), 7.29 (d, J=4.5 Hz, 1H), 5.86 (dd, J=2.8, 1.7Hz, 1H), 4.20 (q, J=7.2 Hz, 2H), 2.65-2.24 (m, 5H), 2.15-1.96 (m, 2H),1.73-1.54 (m, 2H), 1.36-1.29 (m, 3H)

230B. Ethyl 2-(4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)acetate

The reaction mixture of ethyl2-(4-(6-(trifluoromethyl)quinolin-4-yl)cyclohex-3-en-1-yl)acetate (3.0g, 8.26 mmol), ammonium formate (2.08 g, 33.0 mmol) in MeOH (50 mL) waspurged with nitrogen stream for 3 min, followed by addition of Pd—C(0.88 g, 0.41 mmol). The resulting mixture was heated at 85° C. for 2 h.The reaction mixture was cooled down. The reaction mixture was filteredthrough a CELITE® pad and the filter cake was washed with MeOH. Thefiltrate was concentrated in vacuo. The residue was extracted with ethylacetate and washed with saturated NaHCO₃ solution, brine successively.The organic layer was dried over MgSO₄ and the filtrate was concentratedin vacuo to give Intermediate 230B (oil, 2.6 g, 7.12 mmol, 86% yield) asa mixture of cis- and trans-diastereomers. LC-MS Anal. Calc'd forC₂₀H₂₂F₃NO₂ 365.16. found [M+H] 366.2. T_(r)=0.94 min (Method A). ¹H NMR(400 MHz, CHLOROFORM-d) δ: 9.05-8.85 (m, 1H), 8.36 (s, 1H), 8.24 (d,J=8.8 Hz, 1H), 7.88 (dd, J=8.9, 1.7 Hz, 1H), 7.51-7.33 (m, 1H),4.29-4.03 (m, 2H), 3.51-3.23 (m, 1H), 2.61-2.29 (m, 2H), 2.12-1.35 (m,9H), 1.32-1.21 (m, 3H)

230C ethyl2-((1s,4s)-4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)butanoate

To the flask containing THF (15 mL) was added lithium diisopropylamide(2.0 M solution in THF) (7.65 mL, 15.30 mmol) at −78° C., followed byaddition of 1,3-dimethyltetrahydropyrimidin-2(1H)-one (1.29 mL, 10.67mmol) and a solution of ethyl2-(4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)acetate (2.6 g, 7.12mmol) in THF (10 mL) dropwise at −78° C. The resulting mixture turnedinto dark brown solution and stirred at −78° C. for 1 h, then iodoethane(1.14 mL, 14.23 mmol) was added slowly. The reaction mixture was warmedto rt and stirred for 3 h. The reaction was quenched by pouring intowater and extracted with EtOAc. Combined organics was washed with brine,dried with MgSO₄, filtered and the filtrate was concentrated in vacuo.The extract was purified via silica gel flash column chromatography,eluting with 0-20% ethyl acetate in hexane to give the minor isomer andthe major isomer as cis Intermediate 230C (oil, 1.1 g, 2.77 mmol, 39%yield). LC-MS Anal. Calc'd for C₂₂H₂₆F₃NO₂393.19. found [M+H] 394.3.T_(r)=0.97 min (Method A). ¹H NMR (400 MHz, CHLOROFORM-d) δ: 8.97 (d,J=4.6 Hz, 1H), 8.37 (s, 1H), 8.24 (d, J=8.8 Hz, 1H), 7.88 (dd, J=8.8,2.0 Hz, 1H), 7.46 (d, J=4.6 Hz, 1H), 4.20 (q, J=7.2 Hz, 2H), 3.57-3.32(m, 1H), 2.64 (td, J=10.8, 4.0 Hz, 1H), 2.14-1.58 (m, 11H), 1.29 (t,J=7.2 Hz, 3H), 0.95 (t, J=7.4 Hz, 3H).

230D. 2-((1s,4s)-4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)butanoicacid

To the reaction mixture of ethyl2-((1s,4s)-4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)butanoate (1.1g, 2.80 mmol) in THF (20 mL) and MeOH (8 mL) was added lithium hydroxidesolution (2.0 M solution) (13.98 mL, 28.0 mmol). The resulting mixturewas heated at 65° C. over the weekend. The reaction mixture was cooleddown and diluted with water. To the mixture was added 1 N HCl solutionto adjust pH to about 5. White solid crashed out at pH 5-6. Theresulting mixture was extracted with ethyl acetate twice. The organiclayer was separated and washed with brine, dried over MgSO₄. Thefiltrate was concentrated in vacuo to give Intermediate 230D as aracemate (yellow solid, 0.93 g, 2.55 mmol, 91% yield). LC-MS Anal.Calc'd for C₂₀H₂₂F₃NO₂ 365.16. found [M+H] 366.3. T_(r)=0.81 min (MethodA). ¹H NMR (400 MHz, DMSO-d₆) δ: 12.10 (br. s., 1H), 8.99 (d, J=4.6 Hz,1H), 8.57 (s, 1H), 8.23 (d, J=8.8 Hz, 1H), 8.00 (dd, J=8.7, 1.9 Hz, 1H),7.65 (d, J=4.6 Hz, 1H), 3.61 (d, J=10.3 Hz, 1H), 1.96-1.54 (m, 11H),1.49-1.29 (m, 1H), 0.90 (t, J=7.4 Hz, 3H)

230E1-((1s,4s)-4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)propan-1-amine

To a suspension of2-((1s,4s)-4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)butanoic acid(0.58 g, 1.587 mmol) in toluene (15 mL) were added diphenylphosphorylazide (0.40 mL, 1.83 mmol) and triethylamine (0.24 mL, 2.06 mmol). Thereaction mixture turned into clear solution after addition of TEA. Thereaction mixture was heated to 70° C. for 2 h. The reaction was cooledto rt. The reaction mixture was concentrated under reduced pressure. Tothe residue was added THF (15 mL) and 2.0 M lithium hydroxide solution(7.94 mL, 15.87 mmol) and the resulting mixture was stirred at rt for 4h. The reaction mixture was acidified with 1N HCl (white precipitateforms) and extracted with EtOAc to remove DPPA related impurities. Thenthe aqueous layer was basified with 1N NaOH (precipitate forms again)and extracted with EtOAc four times. The organic extracts were combined,dried over MgSO₄ and the filtrate was concentrated in vacuo to givelight yellow oil, dried on high vacuum over night to give Intermediate230E (oil, 0.47 g, 1.397 mmol, 88% yield). LC-MS Anal. Calc'd forC₁₉H₂₃F₃N₂, 336.18. found [M+H] 337.2. T_(r)=0.68 min (Method A). ¹H NMR(400 MHz, CHLOROFORM-d) δ: 8.95 (d, J=4.6 Hz, 1H), 8.38 (s, 1H), 8.24(d, J=8.8 Hz, 1H), 7.88 (dd, J=8.8, 1.8 Hz, 1H), 7.45 (d, J=4.6 Hz, 1H),3.57-3.44 (m, 1H), 2.90 (td, J=8.5, 3.0 Hz, 1H), 2.22-1.20 (m, 13H),1.01 (d, J=15.0 Hz, 3H)

Example 230N-(1-((1s,4s)-4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)propyl)biphenyl-4-carboxamide

To a solution of [1,1′-biphenyl]-4-carboxylic acid (21.2 mg, 0.107 mmol)in DMF (1.5 mL) was added HATU (44 mg, 0.116 mmol). The reaction mixturewas stirred at rt for 10 min, followed by addition of a solution of1-((1s,4s)-4-(6-(trifluoromethyl)quinolin-4-yl)cyclohexyl)propan-1-amine(30 mg, 0.089 mmol) in THF (0.8 mL) and DIPEA (0.03 mL, 0.178 mmol). Thereaction mixture was stirred at rt for 2 h. and was concentrated invacuo. The residue was dissolved in MeOH, filtered, and purified viapreparative HPLC to give a racemic Example 230 (33 mg, 0.063 mmol, 71%yield). LC-MS Anal. Calc'd for C₃₂H₃₁F₃N₂O, 516.24. found [M+H] 517.0.T_(r)=2.02 min (Method B). ¹H NMR (500 MHz, DMSO-d₆) δ: 9.01 (d, J=4.5Hz, 1H), 8.55 (s, 1H), 8.30-8.21 (m, 1H), 8.17 (d, J=9.3 Hz, 1H),8.03-7.91 (m, 3H), 7.79-7.67 (m, 4H), 7.61 (d, J=4.5 Hz, 1H), 7.48 (t,J=7.4 Hz, 2H), 7.43-7.37 (m, 1H), 4.33 (d, J=8.7 Hz, 1H), 4.02-3.49 (m,1H), 1.99-1.33 (m, 11H), 0.90 (t, J=7.0 Hz, 3H)

Example 231a-e (Absolute and Relative Stereochemistry Unknown)4-chloro-N-(1-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohexyl)propyl)benzamide

231A. ethyl 2-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohex-3-enyl)acetate

To the reaction mixture of 2-bromo-6-(difluoromethyl)pyridine (1.55 g,7.45 mmol), ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)acetate(2.52 g, 8.57 mmol) in 1,4-Dioxane (20 mL) was added K₂CO₃ (7.45 mL,22.36 mmol) solution and the resulting mixture was purged with nitrogenstream for 3 min, followed by addition of Pd(Ph₃P)₄ (0.431 g, 0.373mmol) and the reaction mixture was further purged with nitrogen streamand then heated at 110° C. under nitrogen for 20 h. The reaction mixturewas diluted with brine and ethyl acetate. The organic layer wasseparated, dried over MgSO₄. The filtrate was concentrated in vacuo andthe residue was purified via silica gel flash column chromatography,eluting with 0-20% ethyl acetate in hexane to give Intermediate 231A(oil, 2.2 g, 7.45 mmol, 99% yield). LC-MS Anal. Calc'd for C₁₆H₁₉F₂NO₂,295.14. found [M+H] 296.2. T_(r)=1.10 min (Method A). ¹H NMR (400 MHz,METHANOL-d₄) δ: 7.92-7.80 (m, 1H), 7.60 (dd, J=8.0, 0.8 Hz, 1H), 7.47(d, J=7.7 Hz, 1H), 6.71 (dd, J=3.1, 2.0 Hz, 1H), 6.65-6.44 (m, 1H),4.20-4.08 (m, 2H), 2.79-2.65 (m, 1H), 2.56-2.39 (m, 2H), 2.36 (d, J=7.0Hz, 2H), 2.20-1.92 (m, 3H), 1.48 (dtd, J=13.0, 10.6, 5.5 Hz, 1H),1.30-1.22 (m, 3H)

231B. ethyl 2-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohexyl)acetate

The reaction mixture of crude ethyl2-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohex-3-en-1-yl)acetate (2.1 g,7.11 mmol), ammonium formate (1.794 g, 28.4 mmol) in MeOH (40 mL) waspurged with nitrogen stream for 3 min, followed by addition of 5% Pd—C(0.757 g, 0.356 mmol). The resulting mixture was heated at 85° C. for 2h. The reaction mixture was cooled down. The reaction mixture wasfiltered and the filter cake was washed with MeOH. The filtrate wasconcentrated in vacuo. The residue was extracted with ethyl acetate andwashed with saturated NaHCO₃ solution, brine successively. The organiclayer was dried over MgSO₄ and the filtrate was concentrated in vacuo togive Intermediate 231B (oil, 1.8 g, 6.05 mmol, 85% yield) as a mixtureof cis- and trans-diastereomers. LC-MS Anal. Calc'd for C₁₆H₂₁F₂NO₂,297.15 found 298.2 [M+H]. T_(r)=1.09 min (Method A). ¹H NMR (400 MHz,CHLOROFORM-d) δ: 7.75 (t, J=7.8 Hz, 1H), 7.55-7.42 (m, 1H), 7.34-7.23(m, 1H), 6.98 (dd, J=14.0, 7.8 Hz, 1H), 6.80-6.42 (m, 1H), 4.33-4.04 (m,2H), 2.91-2.59 (m, 1H), 2.55-2.36 (m, 2H), 2.34-2.20 (m, 1H), 2.07-1.52(m, 8H), 1.32-1.23 (m, 3H)

231C ethyl 2-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohexyl)butanoate

To the flask containing THF (8 mL) was added lithium diisopropylamide(2.0 M solution in THF) (3.70 mL, 7.40 mmol) at −78° C., followed byaddition of 1,3-dimethyltetrahydropyrimidin-2(1H)-one (0.81 mL, 6.73mmol) and a solution of ethyl2-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohexyl)acetate (1.0 g, 3.36mmol) in THF (10 mL) dropwise at −78° C. The resulting mixture turnedinto brown solution and was stirred at −78° C. for 1 h, then iodoethane(0.54 mL, 6.73 mmol) was added slowly. The reaction mixture was thenstirred at −78° C. for 0.5 h, warmed to rt for 20 h. To the reactionmixture was added more lithium diisopropylamide (2.0 M solution in THF)(3.70 mL, 7.40 mmol) (1.8 mL) at ice bath temperature. The reactionmixture was stirred at ice bath temperature for 2 h. The reaction wasquenched by pouring into water and extracted with EtOAc. Combinedorganics were washed with brine, dried with sodium sulfate, filtered andconcentrated in vacuo. The extract was purified via silica gel flashcolumn chromatography, eluting with 0-16% ethyl acetate in hexane togive Intermediate 231C (oil, 0.365 g, 1.122 mmol, 33% yield). LC-MSAnal. Calc'd for C₁₈H₂₅F₂NO₂, 325.18 found [M+H] 326.3. T_(r)=1.12 min(Method A). ¹H NMR (400 MHz, CHLOROFORM-d) δ: 7.82-7.69 (m, 1H), 7.45(d, J=7.5 Hz, 1H), 7.32 (d, J=7.9 Hz, 0.5H), 7.26-7.21 (m, 0.5H),6.84-6.33 (m, 1H), 4.17 (qd, J=7.1, 5.9 Hz, 2H), 2.90 (dt, J=8.8, 4.4Hz, 0.5H), 2.70 (tt, J=12.2, 3.4 Hz, 0.5H), 2.53-2.36 (m, 0.5H),2.18-2.09 (m, 0.5H), 2.06-1.73 (m, 5H), 1.71-1.57 (m, 4H), 1.55-1.44 (m,1H), 1.27 (dt, J=12.8, 7.2 Hz, 4H), 0.90 (t, J=7.4 Hz, 3H)

231D. 2-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohexyl)butanoic acid

To the reaction mixture of ethyl2-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohexyl)butanoate (0.4 g, 1.229mmol) in THF (6 mL) and MeOH (3 mL) was added LiOH solution (6.15 mL,18.44 mmol) at rt. The reaction mixture was then heated 60° C. overnight. To the reaction mixture was added more THF (4 mL) and LiOHsolution (6.15 mL, 18.44 mmol) and the reaction mixture was heated at60° C. for another 3 days. To the reaction mixture was added 2 N HClsolution to adjust pH to about 5-6 and the resulting mixture wasextracted with ethyl acetate twice. The organic layer was separated anddried over MgSO₄. The filtrate was concentrated in vacuo to giveIntermediate 231D (yellow solid, 0.37 g, 1.22 mmol, 99% yield). LC-MSAnal. Calc'd for C₁₆H₂₁F₂NO₂, 297.15. found [M+H] 298.3, T_(r)=0.96 min(Method A).

¹H NMR (400 MHz, METHANOL-d₄) δ: 7.86 (td, J=7.8, 1.5 Hz, 1H), 7.50-7.37(m, 2H), 6.86-6.47 (m, 1H), 2.99-2.84 (m, 0.5H), 2.72 (tt, J=12.2, 3.4Hz, 0.5H), 2.53-2.37 (m, 0.5H), 2.15-1.42 (m, 10.5H), 1.36-1.12 (m, 1H),0.94 (td, J=7.4, 2.9 Hz, 3H)

231E. 1-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohexyl)propan-1-amine

To a suspension of2-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohexyl)butanoic acid (0.32 g,1.076 mmol) in Toluene (8 mL) were added diphenylphosphoryl azide (0.27mL, 1.24 mmol) and triethylamine (0.17 mL, 1.40 mmol). The reactionmixture in a sealed vial turned into clear solution after addition ofTEA. The reaction mixture was heated to 70° C. for 2.5 h. The reactionmixture was concentrated under reduced pressure. To the residue wasadded THF (10 mL) and 2.0 M lithium hydroxide solution (5.4 mL, 10.76mmol) and the resulting mixture was stirred at rt for 1 h. The reactionmixture was acidified with 1N HCl (white precipitate forms) andextracted with EtOAc to remove DPPA related impurities. Then the aqueouslayer was basified with 1N NaOH (precipitate forms again) and extractedwith EtOAc 3 times. The basic extracts were combined, dried over MgSO₄and the filtrate was concentrated in vacuo to give colorless oil, driedon high vacuum over night to give Intermediate 231E (oil, 95 mg, 0.35mmol, 33% yield). LC-MS Anal. Calc'd for C₁₅H₂₂F₂N₂, 268.17. found [M+H]269.5. T_(r)=0.71 min (Method A).

¹H NMR (400 MHz, METHANOL-d₄) δ: 7.84 (td, J=7.8, 2.2 Hz, 1H), 7.54-7.34(m, 2H), 6.82-6.39 (m, 1H), 2.98 (dt, J=7.6, 3.5 Hz, 0.5H), 2.80-2.64(m, 1H), 2.49 (dt, J=8.1, 4.7 Hz, 0.5H), 2.21-1.17 (m, 11H), 0.96 (q,J=7.4 Hz, 3H)

Example 231 Four Isomers4-chloro-N-(1-(4-(6-(difluoromethyl)pyridin-2-yl)cyclohexyl)propyl)benzamide

To a solution of 4-chlorobenzoic acid (30.1 mg, 0.192 mmol) in DMF (1mL) was added HATU (79 mg, 0.208 mmol). The reaction mixture was stirredat rt for 3 min, followed by addition of a solution of Intermediate 231C(43 mg, 0.160 mmol) in THF (1 mL) and DIPEA (0.1 mL, 0.50 mmol). Thereaction mixture was stirred at rt for 1 h. and was concentrated invacuo. The residue was dissolved in MeOH, filtered, and purified viapreparative HPLC to give a mixture of diastereomers Example 231a.

The isomers were further separated by preparative SFC (Method R) to givefirst eluate Example 231b (11.3 mg, 0.027 mmol, 16.8% yield). LC-MSAnal. Calc'd for C₂₂H₂₅ClF₂N₂O, 406.16. found [M+H] 406.9. T_(r)=2.15min (Method B). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.11 (d, J=9.1 Hz, 1H),7.90 (t, J=7.8 Hz, 1H), 7.82 (d, J=8.3 Hz, 2H), 7.57-7.41 (m, 4H),7.02-6.68 (m, 1H), 4.02 (d, J=9.6 Hz, 1H), 2.86 (br. s., 1H), 2.02-1.82(m, 2H), 1.77-1.26 (m, 9H), 0.82 (t, J=7.2 Hz, 3H).

Second eluate Example 231c (10.5 mg, 0.025 mmol, 15.8% yield). LC-MSAnal. Calc'd for C₂₂H₂₅ClF₂N₂O, 406.16. found [M+H] 407.2. T_(r)=2.28min (Method B). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.09 (d, J=9.1 Hz, 1H),7.92 (t, J=7.8 Hz, 1H), 7.84 (d, J=8.4 Hz, 2H), 7.59-7.38 (m, 4H),7.04-6.73 (m, 1H), 4.03 (d, J=8.1 Hz, 1H), 2.88 (br. s., 1H), 2.06-1.24(m, 11H), 0.83 (t, J=7.2 Hz, 3H)

Third eluate Example 231d (8 mg, 0.019 mmol, 12.0% yield). LC-MS Anal.Calc'd for C₂₂H₂₅ClF₂N₂O, 406.16. found [M+H] 407.0. T_(r)=2.12 min(Method B). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.15 (d, J=9.1 Hz, 1H),7.95-7.77 (m, 3H), 7.52 (d, J=8.3 Hz, 2H), 7.49-7.35 (m, 2H), 7.05-6.59(m, 1H), 3.75 (d, J=9.1 Hz, 1H), 2.79-2.58 (m, 1H), 1.97-1.77 (m, 4H),1.71-1.36 (m, 5H), 1.17 (br. s., 2H), 0.84 (t, J=7.2 Hz, 3H).

Fourth eluate Example 231E (8.9 mg, 0.021 mmol, 13.4% yield). LC-MSAnal. Calc'd for C₂₂H₂₅ClF₂N₂O, 406.16. found [M+H] 406.9. T_(r)=2.12min (Method B). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.15 (d, J=9.0 Hz, 1H),7.96-7.80 (m, 3H), 7.53 (d, J=8.3 Hz, 2H), 7.48 (d, J=7.6 Hz, 1H), 7.43(d, J=7.7 Hz, 1H), 6.99-6.72 (m, 1H), 3.75 (d, J=9.1 Hz, 1H), 2.78-2.58(m, 1H), 1.99-1.77 (m, 4H), 1.70-1.38 (m, 5H), 1.17 (br. s., 2H), 0.84(t, J=7.2 Hz, 3H)

Example 232N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(5-methyl-1,3,4-oxadiazol-2-yl)benzamide

232A.4-bromo-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide

To a solution of 4-bromobenzoic acid (354 mg, 1.762 mmol) in DMF (6 mL)was added HATU (670 mg, 1.762 mmol). The reaction mixture was stirred atrt for 5 min, followed by addition of a solution of(R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethanamine (400 mg,1.469 mmol) in THF (3 mL) and DIPEA (0.77 mL, 4.41 mmol). The reactionmixture was stirred at rt for 3 h. The reaction mixture was diluted withethyl acetate and saturated NaHCO₃ solution. The organic layer wasseparated and washed with brine, dried over MgSO₄. The filtrate wasconcentrated in vacuo. and the residue was purified via silica gel flashcolumn chromatography, eluting with 0-70% ethyl acetate in hexane togive Intermediate 232A (white solid, 0.55 g, 1.208 mmol, 82% yield).LC-MS Anal. Calc'd for C₂₄H₂₄BrFN₂O, 454.1. found [M+H] 455.1, 457.1.T_(r)=0.85 min (Method A). ¹H NMR (400 MHz, CHLOROFORM-d) δ: 8.82 (d,J=4.5 Hz, 1H), 8.12 (dd, J=9.3, 5.7 Hz, 1H), 7.73-7.63 (m, 3H),7.62-7.56 (m, 2H), 7.47 (ddd, J=9.2, 8.0, 2.8 Hz, 1H), 7.42 (d, J=4.5Hz, 1H), 5.85 (d, J=9.3 Hz, 1H), 4.61 (tq, J=9.7, 6.5 Hz, 1H), 3.45-3.17(m, 2.15-1.68 (m, 9H), 1.32 (d, J=6.6 Hz, 3H)

232B.N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

To a solution of4-bromo-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)benzamide(0.55 g, 1.208 mmol) in 1,4-dioxane (20 mL) were added potassium acetate(0.356 g, 3.62 mmol) and bis(pinacolato)diboron (0.368 g, 1.449 mmol).The reaction mixture was purged with nitrogen stream for 3 min, followedby addition of PdCl₂(dppf) (0.088 g, 0.121 mmol). The reaction mixturewas heated at 90° C. over night. The reaction mixture was cooled downand diluted with saturated NaHCO₃ solution and ethyl acetate. Theorganic layer was separated and washed with brine, dried over MgSO₄. Thefiltrate was concentrated in vacuo. to give crude Intermediate 232B asboronic ester and acid mixture (black solid, 0.6 g, 1.208 mmol, 99%yield). LC-MS Anal. Calc'd for C₃₀H₃₆BFN₂O₃, 502.28. found [M+H] 503.5.T_(r)=0.87 min (Method A).

Example 232N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(5-methyl-1,3,4-oxadiazol-2-yl)benzamide

To the reaction mixture of 2-bromo-5-methyl-1,3,4-oxadiazole (15.57 mg,0.096 mmol) and crudeN-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide(40 mg, 0.080 mmol) in dioxane (2 mL) was added Na₂CO₃ (2.0 M solution)(0.12 mL, 0.24 mmol). The reaction mixture was purged with nitrogenstream for 2 min, followed by addition of PdCl₂(dppf) (5.8 mg, 0.0080mmol). The resulting mixture in the sealed tube was heated at 90° C. for16 h. The reaction mixture was diluted with ethyl acetate and sat.NaHCO₃ solution. The organic layer was separated and concentrated invacuo. The residue was dissolved in DMF, filtered, and purified viapreparative HPLC to give Example 232 (17 mg, 0.037 mmol, 46.1% yield).LC-MS Anal. Calc'd for C₂₇H₂₇FN₄O₂ 458.21. found [M+H] 458.9. T_(r)=1.23min (Method I). ¹H NMR (500 MHz, DMSO-d₆) δ: 8.83 (d, J=4.5 Hz, 1H),8.47 (d, J=8.8 Hz, 1H), 8.14-8.00 (m, 5H), 7.97 (d, J=8.8 Hz, 1H), 7.66(t, J=7.4 Hz, 1H), 7.48 (d, J=4.3 Hz, 1H), 4.46 (br. s., 1H), 3.38 (br.s., 1H), 2.59 (s, 3H), 1.96-1.54 (m, 9H), 1.22 (d, J=6.4 Hz, 3H)

Examples 233-253

Examples 233-253 were prepared from Intermediate 40L following theprocedure for Example 47 using the corresponding acid or following theprocedure for Example 231.

Tr Ex. No. Name R (min)^(Method 1*) [M + H]⁺ Example 233N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(pyrazin-2- yl)benzamide

1.47 455.0 Example 234 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(pyrimidin- 5-yl)benzamide

1.20 454.9 Example 235 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(1-methyl- 1H-imidazol-4-yl)benzamide

0.98 457.0 Example 236 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(2- methoxypyrimidin-4-yl)benzamide

1.45 485.1 Example 237 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(6- methylsulfonyl)pyridin-3- yl)benzamide

1.31 532.1 Example 238 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(2- methylthiazol-5-yl)benzamide

1.48 473.9 Example 239 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(5- methoxypyridin-2-yl)benzamide

1.21 483.9 Example 240 4-(2-cyanopyrimidin-5-yl)-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4- yl)cyclohexyl)ethyl)benzamide

1.55 480.4 Example 241 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(2- methoxythiazol-4-yl)benzamide

1.67 489.8 Example 242 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4′-(2- hydroxypropan-2-yl)biphenyl-4- carboxamide

1.56 511.0 Example 243 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(thiazol-4- yl)benzamide

1.51 460.0 Example 244 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(1,3,4- oxadiazol-2-yl)benzamide

1.16 444.9 Example 245 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(6- methoxypyridin-3-yl)benzamide

1.63 484.4 Example 246 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4- morpholinobenzamide

1.29 462.1 Example 247 4-cyclopropyl-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4- yl)cyclohexyl)ethyl)benzamide

1.56 416.9 Example 248 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(1- methylcyclopropyl)benzamide

1.67 431.0 Example 249 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4- (trifluoromethyl)benzamide

1.58 444.9 Example 250 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(oxazol-5- yl)benzamide

1.27 444.0 Example 251 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(1-methyl- 1H-1,2,4-triazol-5-yl)benzamide

1.24 458.3 Example 252 N-((R)-1-((1s,4S)-4-(6- fluoroquinolin-4-yl)cyclohexyl)ethyl)-4-(5- methylthiazol-2-yl)benzamide

1.53 474.1 Example 253 4-(5-cyanothiazol-2-yl)-N-((R)-1-((1s,4S)-4-(6-fluoroquinolin-4- yl)cyclohexyl)ethyl)benzamide

1.61 485.2 *unless otherwise noted

Examples 254-256

Examples 254-256 were prepared from Intermediate 230E following theprocedure for Example 230 using the corresponding acid.

Ex. No. Name R Tr (min)^(Method 1*) [M + H]⁺ Example 2544-chloro-N-(1-((1s,4s)-4-(6- (trifluoromethyl)quinolin-4-yl)cylcohexyl)propyl)benzamide

1.81 474.8 Example 255 4-(1H-pyrrol-1-yl)-N-(1-((1s,4S)-4-(6-(trifloromethyl)quinolin-4- yl)cyclohexyl)propyl)benzamide

1.89 506.1 Example 256 6-methoxy-N-(1-((1s,4S)-4-(6-(trifluoromethyl)quinolin-4- yl)cylcohexyl)propyl)nicotinamide

1.56 471.9 *unless otherwise noted

Examples 257-263

Examples 257-263 were prepared from Intermediate 164J following theprocedure for Example 164 using the corresponding acid.

Ex. No. Stereochemistry Name R Tr (min)^(Method 1*) [M + H]⁺ Example 257Diastereomer mixture N-(1-(4-(6- fluoroquinolin-4-yl)cyclohexyl)propyl)-4- (thiazol-2-yl)benzamide

1.53- 1.56 474.3 Example 258 Diastereomer mixture N-(1-(4-(6-fluoroquinolin-4- yl)cyclohexyl)propyl)bi- phenyl-4-carboxamide

1.79- 1.83 467.0 Example 259 Isomer 1 Homochiral, absolute and realtivestereochemistry unknown N-(1-(4-(6- fluoroquinolin-4-yl)cyclohexyl)propyl)bi- phenyl-4-carboxamide

1.83 466.9 Example 260 Isomer 2 Homochiral, absolute and realtivestereochemistry unknown N-(1-(4-(6- fluoroquinolin-4-yl)cylcohexyl)propyl)bi- phenyl-4-carboxamide

1.93 467.1 Example 261 Isomer 3 Homochiral, absolute and realtivestereochemistry unknown N-(1-(4-(6- fluoroquinolin-4-yl)cyclohexyl)propyl)bi- phenyl-4-carboxamide

1.90 467.1 Example 262 Isomer 4 Homochiral, absolute and realtivestereochemistry unknown N-(1-(4-(6- fluoroquinolin-4-yl)cyclohexyl)propyl)bi- phenyl-4-carboxamide

1.85 467.4 Example 263 Diastereomer mixture N-(1-(4-(6-fluoroquinolin-4- yl)cyclohexyl)propyl)-6- methoxynicotinamide

1.35- 1.39 422.3 *unless otherwise noted

BIOLOGICAL EXAMPLES

Assessment of inhibitor activity in HeLa cell-based indoleamine2,3-dioxygenase (IDO) assay.

HeLa (ATCC® CCL-2) cells were obtained from the ATCC® and cultured inDulbecco's Modified Eagle Medium supplemented with 4.5 g/L glucose, 4.5g/L L-glutamine and 4.5 g/L sodium pyruvate (#10-013-CV, Corning), 2 mML-alanyl-L-glutamine dipeptide (#35050-061, Gibco), 100 U/mL penicillin,100 μg/mL streptomycin (#SV30010, HyClone) and 10% fetal bovine serum(#SH30071.03 HyClone). Cells were maintained in a humidified incubatorat 37° C. in 5% CO₂.

IDO activity was assessed as a function of kynurenine production asfollows: HeLa cells were seeded in a 96-well culture plate at a densityof 5,000 cells/well and allowed to equilibrate overnight. After 24hours, the media was aspirated and replaced with media containing IFNγ(#285-IF/CF, R&D Systems) at a final concentration of 25 ng/mL. A serialdilution of each test compound was added to the cells in a total volumeof 200 μL of culture medium. After a further 48 hour incubation, 170 μLof supernatant was transferred from each well to a fresh 96-well plate.12.1 μL of 6.1N trichloroacetic acid (#T0699, Sigma-Aldrich) was addedto each well and mixed, followed by incubation at 65° C. for 20 minutesto hydrolyze N-formylkynurenine, the product of indoleamine2,3-dioxygenase, to kynurenine. The reaction mixture was thencentrifuged for 10 mins at 500×g to sediment the precipitate. 100 μL ofthe supernatant was transferred from each well to a fresh 96-well plate.100 μl of 2% (w/v) p-dimethylaminobenzaldehyde (#15647-7, Sigma-Aldrich)in acetic acid (#A6283, Sigma-Aldrich) was added to each well mixed andincubated at room temperature for 20 mins. Kynurenine concentrationswere determined by measuring absorbance at 480 nm and calibratingagainst an L-kynurenine (#K8625, Sigma-Aldrich) standard curve using aSPECTRAMAX® M2e microplate reader (Molecular Devices). The percentageactivity at each inhibitor concentration was determined and IC₅₀ valuesassessed using nonlinear regression.

Activity for compounds described herein is provided in FIG. 1, whereinpotency levels are provided as follows: (Potency: IDO IC₅₀: A<0.1 μM;B<1 μM; C<10 μM)

Evaluation of Biological Activity

Exemplary compounds were tested for inhibition of IDO activity.Experimental procedures and results are provided below.

HEK293 cells were transfected with a pCDNA-based mammalian expressionvector harboring human IDO1 cDNA (NM 002164.2) by electroporation. Theywere cultured in medium (DMEM with 10% FBS) containing 1 mg/ml G418 fortwo weeks. Clones of HEK293 cells that stably expressed human IDO1protein were selected and expanded for IDO inhibition assay.

The human IDO1/HEK293 cells were seeded at 10,000 cells per 504 per wellwith RPMI/phenol red free media contains 10% FBS in a 384-well blackwall clear bottom tissue culture plate (Matrix Technologies LLC) 100 nLof certain concentration of compound was then added to each well usingECHO liquid handling systems. The cells were incubated for 20 hours in37° C. incubator with 5% CO₂.

The compound treatments were stopped by adding trichloroacetic acid(Sigma-Aldrich) to a final concentration at 0.2%. The cell plate wasfurther incubated at 50° C. for 30 minute. The equal volume supernatant(20 μL) and 0.2% (w/v) Ehrlich reagent (4-dimethylaminobenzaldehyde,Sigma-Aldrich) in glacial acetic acid were mixed in a new clear bottom384-well plate. This plate was then incubated at room temperature for 30minute. The absorbance at 490 nm was measured on Envision plate reader.

Compound IC₅₀ values were calculated using the counts of 500 nM of areference standard treatment as one hundred percent inhibition, andcounts of no compound but DMSO treatment as zero percent inhibition.

Activity for compounds described herein is provided in FIG. 1, whereinpotency levels are provided as follows: (Potency: IDO IC₅₀: A<0.05 μM;B<0.25 μM; C<2 μM)

Results of the IDO assays are shown in the table below.

HEK Human IDO-1 Example Bio Activity No. A < 0.05, b < 0.250, c < 2.0 40 C  41 B  42 C  43 C  44 A  45 C  46 C  47 A  48 A  49 A  50 A  51 A 52 A  53 A  54 A  55 A  56 A  57 A  59 A  60 A  61 A  66 A  67 A  68 A 69 A  70 A  71 A  72 C  75 B  76 C  77 B  78 C  79 A  80 C  81 C  84 C 86 B  87 B  88 C  89 B  90 C  91 B  92 C  93 C  97 B 119 A 120 A 121 B122 B 123 B 124 A 125 B 130 B 131 B 139 C 140 ? 144 C 145 C 146 C 147 C148 C 149 B 157a B 157b A 157c C 157d C 157e C 158a A 158b A 158c A 158dA 158e C 159a A 159b B 159c A 159d A 159e C 160a A 160b C 160c C 160d B160e A 161a C 161b A 161c B 161e C 163 A 164a B 164b A 164c A 164d A165a C 165b A 165c A 165d A 176 A 178 B 194 A 195 A 196 A 197 B 198 A199 C 200 B 201 B 202 A 203a C 203b B 203c A 203d A 207 A 208 B 209 B210 B 211 B 212 C 213 A 214 B 215 C 216 B 217 C 218 C 219 B 220 B 221 A222 C 223 A 224 B 225 Nt 226 C 227 C 228 C 229 Nt 230 A 231a B 231b C231c C 231d C 231e A 232 B 233 A 234 A 235 A 236 A 237 C 238 A 239 A 240C 241 A 242 B 243 A 244 A 245 A 246 B 247 A 248 A 249 A 250 A 251 B 252A 253 A 254 A 255 A 256 A 257 Nt 258 A 259 260 261 262 263

Particular embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention.Upon reading the foregoing, description, variations of the disclosedembodiments may become apparent to individuals working in the art, andit is expected that those skilled artisans may employ such variations asappropriate. Accordingly, it is intended that the invention be practicedotherwise than as specifically described herein, and that the inventionincludes all modifications and equivalents of the subject matter recitedin the claims appended hereto as permitted by applicable law. Moreover,any combination of the above-described elements in all possiblevariations thereof is encompassed by the invention unless otherwiseindicated herein or otherwise clearly contradicted by context.

All publications, patent applications, accession numbers, and otherreferences cited in this specification are herein incorporated byreference as if each individual publication or patent application werespecifically and individually indicated to be incorporated by reference.

1. A compound having the formula (I):

or a pharmaceutically acceptable salt, hydrate or solvate thereof,wherein, the subscript n is 1 or 0; A is —C(O)—, —NH—, —SO₂—, —CH₂—, or—CHR³—; B is a bond, —C(O)—, —NH—, —CH₂—, or —CHR³—; T is a bond, —CH₂—,—NH—, —O—, —OCH₂—, —C(O)CH₂—, or —CR³R⁴—; wherein when A is —NH— and Bis —C(O)—, then T is other than —C(R³)(R⁴)—; D is N or C(R⁵); E is N orC(R⁶); V is a bond, —O—, or —C(R^(5a))₂; G is an optionally substitutedaryl, optionally substituted heteroaryl, or an optionally substituted 9-or 10-membered fused bicyclic heteroaryl; J¹ is CH, N or C(R²), when R²is attached to the ring vertex identified as J¹; R¹ and R² areindependently hydrogen, halogen, optionally substituted C₁-C₄ haloalkyl,optionally substituted C₃-C₆ cycloalkyl, optionally substituted 3- to6-membered cycloheteroalkyl, optionally substituted phenyl, optionallysubstituted heteroaryl, optionally substituted C₁-C₄ alkyl, optionallysubstituted C₁-C₄ alkoxy, CN, SO₂NH₂, NHSO₂CH₃, NHSO₂CF₃, OCF₃, SO₂CH₃,SO₂CF₃, or CONH₂, and when R¹ and R² are on adjacent vertices of aphenyl ring they may be joined together to form a 5- or 6-memberedcycloheteroalkyl ring having one or two ring vertices independentlyselected from O, N and S, wherein said cycloheteroalkyl ring isoptionally substituted with from one to three members selected fromfluoro and C₁-C₃ alkyl; R³ and R⁴ are independently hydrogen, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ haloalkyl,fluorine, OH, CN, CO₂H, C(O)NH₂, N(R^(5a))₂, optionally substituted—O—C₁-C₆ alkyl, —(CR⁵R⁵)_(m)—OH, —(CR⁵R⁵)_(m)—CO₂H,—(CR⁵R⁵)_(m)—C(O)NH₂, —(CR⁵R⁵)_(m)—C(O)NHR^(5a), —(CR⁵R⁵)_(m)N(R^(5a))₂,—NH(CR⁵R⁵)_(m)CO₂H or —NH(CR⁵R⁵)_(m)—C(O)NH₂; each R⁵ is independentlyH, F, OH, optionally substituted C₁-C₆ alkyl or optionally substituted—O—C₁-C₆ alkyl; each R^(5a) is independently H, or optionallysubstituted C₁-C₆ alkyl; R⁶ is H, OH, F, optionally substituted C₁-C₆alkyl, optionally substituted —O—C₁-C₆ alkyl, or —N(R^(5a))₂; and each mis independently 1, 2, or
 3. 2. A compound of claim 1, having theformula:


3. A compound of claim 2, having the formula:


4. A compound of claim 3, having the formula:


5. A compound of claim 4, having the formula:


6. A compound of claim 4, having the formula:


7. A compound of claim 4, having the formula:


8. A compound of claim 4, having the formula:


9. A compound of claim 4, having the formula:


10. A compound of claim 9, having the formula:


11. A compound of claim 9, having the formula:


12. A compound of claim 9, having the formula:


13. A compound of claim 9, having the formula:


14. A compound of claim 1, having the formula:


15. A compound of claim 1, having the formula:


16. A compound of claim 1, having the formula:


17. A compound of claim 1, having the formula:


18. A compound of claim 17, having the formula:


19. A compound of claim 1, having the formula:


20. A compound of claim 1, having the formula:


21. A compound of claim 1, having the formula:


22. A compound of claim 1, having the formula:


23. A compound of claim 1 as provided in the examples.
 24. A compound ofclaim 1, having the formula:


25. A pharmaceutical composition comprising a compound of claim 1 and apharmaceutically acceptable excipient.
 26. A method of treating adisease, disorder or condition, mediated at least in part by IDO, saidmethod comprising administering an effective amount of a compound ofclaim 1, to a subject in need thereof.
 27. A method of claim 26, whereinsaid disease, disorder or condition is cancer.
 28. A method of claim 27,wherein said cancer is a cancer of the prostate, colon, rectum,pancreas, cervix, stomach, endometrium, brain, liver, bladder, ovary,testis, head, neck, skin (including melanoma and basal carcinoma),mesothelial lining, white blood cell (including lymphoma and leukemia),esophagus, breast, muscle, connective tissue, lung (including small-celllung carcinoma and non-small-cell carcinoma), adrenal gland, thyroid,kidney, or bone; or is glioblastoma, mesothelioma, renal cell carcinoma,gastric carcinoma, sarcoma (including Kaposi's sarcoma),choriocarcinoma, cutaneous basocellular carcinoma, or testicularseminoma.
 29. A method of claim 27, wherein said cancer is selected fromthe group consisting of melanoma, colon cancer, pancreatic cancer,breast cancer, prostate cancer, lung cancer, leukemia, a brain tumor,lymphoma, ovarian cancer, and Kaposi's sarcoma.
 30. A combinationcomprising a compound of claim 1 and at least one additional therapeuticagent.
 31. A combination of claim 30, wherein the at least oneadditional therapeutic agent is a chemotherapeutic agent, an immune-and/or inflammation-modulating agent, an anti-hypercholesterolemiaagent, or an anti-infective agent.
 32. A combination of claim 30,wherein the at least one additional therapeutic agent is an immunecheckpoint inhibitor.
 33. A method of treating cancer in a subject, saidmethod comprising administering to said subject an effective amount of acompound of claim 1 and an immune checkpoint inhibitor.
 34. Acombination or method of claim 32, wherein said immune checkpointinhibitor is selected from the group consisting of ipilimumab, nivolumaband pembrolizumab.